Compounds, methods and formulations for the oral delivery of a glucagon-like peptide (glp-1) compound or a melanocortin-4 receptor (mic4) agonist peptide

ABSTRACT

The present invention relates to novel compounds, methods, and formulations useful for the oral delivery of a GLP-1 compound or an MC4 agonist peptide.

BACKGROUND OF THE INVENTION

Conventional means for delivering active agents are often severelylimited by biological, chemical, and physical barriers. Typically, thesebarriers are imposed by the environment through which delivery occurs,the environment of the target for delivery, or the target itself.Biologically or chemically active agents are particularly vulnerable tosuch barriers. In the delivery to animals of biologically active orchemically active pharmacological and therapeutic agents, physical andchemical barriers are imposed by the body. Examples of physical barriersare the skin and various organ membranes that must be traversed beforereaching a target, and examples of chemical barriers include, but arenot limited to, variations in pH, lipid bilayers, and degrading enzymes.

These barriers are of particular significance in the design of oraldelivery systems. Oral delivery of many biologically or chemicallyactive agents would be the route of choice for administration to animalsif not for biological, chemical, and physical barriers such as varyingpH in the gastrointestinal (GI) tract, powerful digestive enzymes, andactive agent impermeable gastrointestinal membranes. Among the numerousagents which are not typically amenable to oral administration arebiologically or chemically active peptides, such as calcitonin andinsulin; polysaccharides, and in particular mucopolysaccharidesincluding, but not limited to, heparin; heparinoids; antibiotics; andother organic substances. These agents are rapidly rendered ineffectiveor are destroyed in the gastrointestinal tract by acid hydrolysis,enzymes, or the like.

Earlier methods for orally administering vulnerable pharmacologicalagents have relied on the co-administration of excipients or enhancers(e.g., resorcinols and non-ionic surfactants such as polyoxyethyleneoleyl ether and n-hexadecylpolyethylene ether) to increase artificiallythe permeability of the intestinal walls, as well as theco-administration of enzyme inhibitors (e.g., pancreatic trypsininhibitors, diisopropylfluorophosphate) to inhibit enzymaticdegradation.

Liposomes have also been described as drug delivery systems for insulinand heparin. See, for example, U.S. Pat. No. 4,239,754; Patel et al(1976), FEBS Letters, Vol 62, pg. 60, and Hashimoto et al. (1970),Endocrinology Japan, Vol, 26, pg. 337.

However, broad spectrum use of such drug delivery systems is precludedbecause: (1) the systems require toxic amounts of excipients, enhancersor inhibitors; (2) suitable low molecular weight cargos, i.e. activeagents, are not available; (3) they exhibit poor stability andinadequate shelf life; (4) the systems are difficult to manufacture; (5)the systems fail to protect the active agent (cargo); (6) the systemsadversely alter the active agent; or (7) the systems fail to allow orpromote absorption of the active agent.

More recently, microspheres of artificial polymers of mixed amino acids(proteinoids) have been used to deliver pharmaceuticals. For example,U.S. Pat. No. 4,925,673 describes drug-containing proteinoid microspherecarriers as well as methods for their preparation and use. Theseproteinoid microspheres are useful for the delivery of a number ofactive agents.

Delivery agent molecules have also been disclosed in U.S. Pat. Nos.5,541,155; 5,693,338; 5,976,569; 5,643,957; 5,955,503; 6,100,298;5,650,386; 5,866,536; 5,965,121; 5,989,539; 6,001,347; 6,071,510;5,820,881; and 6,242,495; see also WO 02/02509; WO 01/51454; WO01/44199; WO 01/32130; WO 00/59863; WO 00/50386; WO 00/47188; and WO00/40203.

BRIEF SUMMARY OF THE INVENTION

The present invention relates to a compound of formula I:

wherein

R¹ and R² are each independently H, OH, cyano, C₁-C₆ alkyl, C₁-C₆alkoxy, CF₃, halo or NR⁴R^(4′);

R³ is H, C₁-C₆ alkyl;

R⁴ is H, COR⁵, SO₂R⁶, or C₁-C₆ alkyl;

R^(4′) is H or C₁-C₆ alkyl;

R⁵ is H or C₁-C₆ alkyl;

R⁶ is H or C₁-C₆ alkyl;

X is a 5 membered aromatic heterocycle that is optionally substitutedwith C1-C4 alkyl; wherein said heterocycle contains at least two orthree heteroatoms selected from N, S and O wherein at least oneheteroatom must be N and wherein said heterocycle may not be1,3,4-oxadiazole;

n is 2, 3, 4, 5, 6 or 7;

or a pharmaceutical salt thereof.

The present invention further relates to a compound of formula I whereinR³ is H. This compound is hereafter referred to as a compound of formulaII.

The present invention also relates to a pharmaceutical compositioncontaining a compound of formula II, or a pharmaceutical salt thereof,and a pharmaceutical carrier.

The present invention also relates to a pharmaceutical compositioncontaining a compound of formula II, or a pharmaceutical salt thereof,and a GLP-1 compound.

The present invention also relates to a pharmaceutical compositioncontaining a compound of formula II, or a pharmaceutical salt thereof,and a MC4 agonist peptide.

DETAILED DESCRIPTION OF THE INVENTION

Reference hereafter to “a compound of formula I” or “compound of formulaII” includes the pharmaceutical salts thereof.

For the purposes of the present invention, as disclosed and claimedherein, the following terms are defined below.

The term “halo” refers to fluoro, chloro, bromo and iodo. The term“C₁-C₆ alkyl” represents a straight, branched or cyclic hydrocarbonmoiety having from one to six carbon atoms, e.g., methyl, ethyl,n-propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, t-butyl,cyclobutyl, pentyl, cyclopentyl, hexyl, cyclohexyl and the like.Moieties such as a cyclobutylmethylene are also included within thescope of a C₁-C₆ alkyl group. The term “C₁-C₄ alkyl” refers specificallyto methyl, ethyl, n-propyl, isopropyl, cyclopropyl, cyclopropylmethyl,n-butyl, isobutyl, sec-butyl, t-butyl and cyclobutyl. A “C₁-C₆ alkoxy”group is a C₁-C₆ alkyl moiety connected through an oxy linkage.

The term “pharmaceutical” when used herein as an adjective meanssubstantially non-deleterious to the recipient patient.

The term “patient” includes humans and non-human animals such ascompanion animals (dogs, cats, horses and the like). The preferredpatient of treatment is a human.

The term “GLP-1 compound” as used herein refers to one or more naturallyoccurring GLP-1 polypeptides (GLP-1(7-37)OH and GLP-1(7-36)NH₂), GLP-1fragments, GLP-1 analogs, GLP-1 derivatives of naturally occurring GLP-1polypeptides, GLP-1 fragments, or GLP-1 analogs, and Exendin-3 andExendin-4 that have the ability to bind to the GLP-1 receptor andinitiate a signal transduction pathway resulting in insulinotropicactivity as described in PCT Publication Number WO 03/072195(Application Number PCT/US03/03111); herein incorporated by reference.

The term “MC4 agonist peptide” as used herein refers to thepharmaceutically useful peptides disclosed in PCT Patent Application No.PCT/US04/16625, filed Jun. 17, 2004 (peptides of formula I, II and IIIas disclosed therein).

The compound of formula II is useful for increasing the oralbioavailability of an active agent, i.e., a GLP-1 compound or an MC4agonist peptide, when said compound is mixed with the active agent toform a combination composition. Said combination is one embodiment ofthe present invention. The compositions of the present inventioncomprise a compound of formula II, that is, a delivery agent (a formulaII compound), and a GLP-1 compound or an MC4 agonist peptide.

The present invention is particularly advantageous for delivering aGLP-1 compound or an MC4 agonist peptide (active agent) that wouldotherwise be destroyed or rendered less effective by conditionsencountered before the active agent reaches its target zone (i.e. thearea in which the active agent of the delivery composition is to bereleased) and within the body of the animal to which it is administered.The compositions comprising one or more compounds of formula II(preferably and most typically one) and an active agent have utility inthe delivery of said active agent to selected biological systems and inan increased or improved bioavailability of the active agent compared toadministration of the active agent without the delivery agent. Deliverycan be improved by delivering more active agent over a period of time,or in delivering active agent in a particular time period (such as toeffect quicker or delayed delivery) or over a period of time (such assustained delivery).

Preferred Compounds (Embodiments) of the Invention

Certain compounds of the invention are particularly interesting and arepreferred. The following listing sets out several groups of preferredcompounds. It will be understood that each of the listings may becombined with other listings to create additional groups of preferredcompounds.

n is 2, 3, 4 or 5;

R¹ and R² are each independently H, OH, OCH₃ CH₃, CF₃, Cl, or Br;

R¹ and R² are each independently H, OH, OCH₃ CH₃ or CF₃;

R¹ and R² are each independently H, OH, OCH₃ or NH₂;

R³ is H;

R⁴ is H;

R⁴ is COR⁵ and R⁵ is CH₃;

R⁴ is SO₂R⁶ and R⁶ is CH₃;

R^(4′) is H;

R⁶ is C₁-C₆ alkyl;

X is

and the phenyl substituent is attached at either carbon atom number 4 or5 and the alkanoic acid is attached at carbon atom number 2;

X is

and the phenyl substituent is attached at either carbon atom number 2 or4 and the alkanoic acid is attached at nitrogen atom number 1;

X is

and the phenyl substituent is attached at carbon atom number 3 and thealkanoic acid is attached at nitrogen atom number 1;

X is

and the phenyl substituent is attached at carbon atom number 3 and thealkanoic acid is attached at carbon atom number 5;

X is

and the phenyl substituent is attached at carbon atom number 3 and thealkanoic acid is attached at carbon atom number 4;

X is

and the phenyl substituent is attached at carbon atom number 5 and thealkanoic acid is attached at carbon atom number 3;

X is

optionally substituted with methyl at carbon atom number 5 where thephenyl substituent is attached at carbon atom number 4 and the alkanoicacid chain is attached at carbon atom number 2;

X is

optionally substituted with methyl at carbon atom number 4 where thephenyl substituent is attached at carbon atom number 2 and the alkanoicacid chain is attached at carbon atom number 5;

X is

and the phenyl substituent is attached at carbon atom number 2 and thealkanoic acid chain is attached at carbon atom number 5;

X is

and the phenyl substituent is attached at carbon atom number 4 and thealkanoic acid chain is attached at carbon atom number 2;

X is

and the phenyl substituent is attached at carbon atom number 5 and thealkanoic acid is attached at carbon atom number 2;

X is

and the phenyl substituent is attached at carbon atom number 4 and thealkanoic acid is attached at carbon atom number 2;

X is

and the phenyl substituent is attached at carbon atom number 5 and thealkanoic acid is attached at carbon atom number 2;

X is

and the phenyl substituent is attached at carbon atom number 2 and thealkanoic acid is attached at carbon atom number 4;

X is

optionally substituted at nitrogen atom number 1 with methyl;

X is

X is

and the phenyl substituent is attached at carbon atom number 4 and thealkanoic acid is attached at nitrogen atom number 1;

X is

and the phenyl substituent is attached at carbon atom number 5 and thealkanoic acid is attached at nitrogen atom number 1;

X is

and the phenyl substituent is attached at carbon atom number 3 and thealkanoic acid is attached at carbon atom number 5;

X is

and the phenyl substituent is attached at carbon atom number 5 and thealkanoic acid is attached at carbon atom number 3;

X is

and the phenyl substituent is attached at carbon atom number 5 and thealkanoic acid is attached at carbon atom number 2.

PREPARATIONS AND EXAMPLES

All non-aqueous reactions are performed under a dry atmosphere ofnitrogen unless otherwise specified. Commercial grade reagents andanhydrous solvents are used as received from vendors and no attempts aremade to purify or dry these components further. Removal of solventsunder reduced pressure is accomplished with a Buchi rotary evaporator atapproximately 28 mm Hg pressure using a Teflon-lined KNF vacuum pump.Thin layer chromatography is performed using 1″×3″ Analtech No. 02521,Whatman No. MK6F or EM Science (Merck) No. 5719-2 silica gel plates withfluorescent indicator. Visualization of TLC plates is made byobservation with either short wave UV light, 10% phosphomolybdic acid inethanol or in iodine vapors. Flash column chromatography is carried outusing Kieselgel silica gel 60. Proton NMR spectra are obtained on aBruker AC 300 MHz Nuclear Magnetic Resonance Spectrometer and arereported in ppm δ values, using tetramethylsilane as an internalreference. Melting points are obtained using an Electrothermal meltingpoint apparatus and are uncorrected. CI Mass spectroscopic analyses areperformed on a Shimadzu QP-5000 GC/Mass Spectrometer (methane) by directinjection. API Mass spectroscopic analyses are performed on a FinneganLCQ Duo Ion Trap or a PESciex API 150EX mass spectrometer, using electrospray ionization (ESI) or atmospheric pressure chemical ionization(APCI). HPLC analyses are conducted using a Waters Symmetry C18, 5 um,WAT046980, 3.9×150 mm column. The elution system consisted of 90:10(0.1% TFA in H₂O)/(0.1% TFA in CH₃CN) gradient elution to 10:90 (0.1%TFA in H₂O)/(0.1% TFA in CH₃CN) over 20 min, followed by 10:90 (0.1% TFAin H₂O)/(0.1% TFA in CH₃CN) isocratic elution for 10 min, followed by90:10 (0.1% TFA in H₂O)/(0.1% TFA in CH₃CN) isocratic elution for 10min. The flow rate is 1 mL/min. UV Detection is performed at both 214and 254 nm.

Preparation 1 Octanedioic Acid Methyl Ester 2-Oxo-2-phenylethyl Ester

Add a solution of sodium bicarbonate (2.12 g, 25.2 mmol) in water (10mL) to a solution of suberic acid monomethyl ester (4.75 g, 25.2 mmol)in methanol (50 mL) at room temperature and stir the mixture for 30minutes. Remove the solvent under reduced pressure and add the residueto a solution of 2-bromoacetophenone (5.0 g, 25.1 mmol) in acetone (150mL) at room temperature under nitrogen. Heat the mixture at reflux for10 hours and then remove the solvent under reduced pressure. Dilute theresidue with diethyl ether (300 mL), stir for 20 minutes, filter througha short silica gel column, and wash with diethyl ether (2×50 mL). Removethe solvent under reduced pressure to provide octanedioic acid methylester 2-oxo-2-phenylethyl ester (6.9 g, 90%).

Example 1 7-(4-Phenyloxazol-2-yl)heptanoic Acid Methyl Ester

Heat a mixture of octanedioic acid methyl ester 2-oxo-2-phenylethylester (6.93 g, 22.6 mmol), acetamide (6.75 g, 114 mmol) and borontrifluoride diethyl etherate (3.0 mL, 23.7 mmol) at 135-140° C. undernitrogen for 4 hours. Cool the mixture, dilute with saturated NaHCO₃solution (100 mL), and extract with ethyl acetate (250 mL). Wash theorganic extract with 100 mL of saturated aqueous sodium chloride (brine)and dry over sodium sulfate. Remove the solvent under reduced pressureand purify the residue by flash column chromatography on silica gel,eluting with hexanes/ethyl acetate (85:15), to provide7-(4-phenyloxazol-2-yl)heptanoic acid methyl ester (5.7 g, 88%).

Example 2 7-(4-Phenyloxazol-2-yl)heptanoic Acid

Add solution of sodium hydroxide (1.60 g, 40.0 mmol) in water (30 mL) toa solution of 7-(4-phenyloxazol-2-yl)heptanoic acid methyl ester (5.75g, 20.0 mmol) in methanol (40 mL) at room temperature and heat themixture at 40° C. for 2 hours. Adjust the pH of the mixture to 2 with 1N HCl and extract with ethyl acetate (600 mL). Wash the organic extractwith water (3×150 mL), dry over sodium sulfate and remove the solventunder reduced pressure. Triturate the residue with hexanes/ethyl acetateand collect the solids by filtration to provide7-(4-phenyloxazol-2-yl)heptanoic acid (5.01 g, 91%): APCI mass spectrumm/z 272 [C₁₆H₁₉NO₃—H]⁻.

Prepare Examples 3-30, compounds of formula II(a) listed in Table 1below, by the same process as in the preparation of Example 2.

TABLE 1 II(a)

Compounds of formula II(a) R¹ (position on R² (position on Example ring)ring) n mass spectrum m/z  3 OCH₃ (1) H 3 261 [C₁₄H₁₅NO₄]⁺  4 OCH₃ (1) H4 276 [C₁₅H₁₇NO₄ + H]⁺  5 OCH₃ (1) H 5 290 [C₁₆H₁₉NO₄ + H]⁺  6 OH (1) H4 260 [C₁₄H₁₅NO₄ − H]⁻  7 OH (1) OCH₃ (3) 4 290 [C₁₅H₁₇NO₅ − H]⁻  8 OCH₃(1) H 6 302 [C₁₇H₂₁NO₄ − H]⁻  9 H OCH₃ (3) 6 302 [C₁₇H₂₁NO₄ − H]⁻  10 OH(1) H 6 288 [C₁₆H₁₉NO₄ − H]⁻  11 OH (1) OCH₃ (3) 6 318 [C₁₇H₂₁NO₅ − H]⁻ 12 OH (1) Cl (4) 6 322 [C₁₆H₁₈ClNO₄ − H]⁻  13 OH (1) H 3 246 [C₁₃H₁₃NO₄− H]⁻  14 OH (1) F (3) 4 280 [C₁₄H₁₄FNO₄ + H]⁺  15 OH (1) H 5 276[C₁₅H₁₇NO₄ + H]⁺  16 OCH₃ (1) H 2 248 [C₁₃H₁₃NO₄ + H]⁺  17 OH (1) H 2234 [C₁₂H₁₁NO₄ + H]⁺  18 OH (1) OCH₃ (3) 3 276 [C₁₄H₁₅NO₅ − H]⁻  19 H H4 246 [C₁₄H₁₅NO₃ + H]⁺  20 F (1) H 4 264 [C₁₄H₁₄FNO₃ + H]⁺  21 OCH₃ (2)H 4 276 [C₁₅H₁₇NO₄ + H]⁺  22 OH (2) H 4 262 [C₁₄H₁₅NO₄ + H]⁺  23 OH (1)OH (3) 4 278 [C₁₄H₁₅NO₅ + H]⁺  24 OH (1) OCH₃ (3) 5 304 [C₁₃H₁₃NO₄ − H]⁻ 25 OH (1) OH (5) 4 278 [C₁₄H₁₅NO₅ + H]⁺  26 OH (1) F (4) 4 278[C₁₄H₁₄FNO₄ − H]⁻  27 OH (1) CN (4) 4 285 [C₁₅H₁₄N₂O₄ − H]⁻  28 OH (1)CN (3) 4 287 [C₁₅H₁₄N₂O₄ + H]⁺  29 OH (1) Br (3) 4 278 [C₁₄H₁₄BrNO₄H]⁺ 30 OH (1) OCH₃ (4) 4 290 [C₁₅H₁₇FNO₅ − H]⁻ 174 OH (1) Cl (3) 4 294[C₁₄H₁₄ClNO₄ − H]⁻ 175 OH (1) Br (4) 4 339 [C₁₄H₁₄BrNO₄ − H]⁻ 176 OH (1)CH₃ (3) 4 276 [C₁₅H₁₇N₂O₄ + H]⁺ 177 OH (1) CH₃ (4) 4 276 [C₁₅H₁₇N₂O₄ +H]⁺ 178 OH (1) N(CH₃)₂ (3) 4 303 [C₁₆H₂₀N₂O₄ − H]⁻ 179 OH (1) NHSO₂CH₃(4) 4 355 [C₁₅H₁₈N₂O₆S + H]⁺ 180 OH (1) NHSO₂CH₃ (3) 4 355[C₁₅H₁₈N₂O₆S + H]⁺

Preparation 2 (6-Bromohexyloxy)-tert-butyldimethylsilane

Add a solution of tert-butyldimethylsilyl chloride (5.0 g, 33.1 mmol) indimethylforamide (DMF) (70 mL) dropwise over 15 minutes to a solution of6-bromohexanol (5.0 g, 27.6 mmol) and imidazole (4.7 g, 69 mmol) in DMF(80 mL) at 0° C. under nitrogen protection and stir the mixture foranother 3.5 hours. Dilute the mixture with water (400 mL) and extractwith diethyl ether (3×150 mL). Dry the combined organic extracts oversodium sulfate and remove the solvent under reduced pressure. Purify thecrude product by flash column chromatography on silica gel, eluting withethyl acetate/hexanes (1:19), to provide(6-bromohexyloxy)-tert-butyldimethylsilane (8.05 g, 98%).

Preparation 3 7-[4-(4-Methoxyphenyl)thiazol-2-yl]heptanal

Add a solution of thioacetamide (2.65 g, 34.9 mmol) in acetone (100 mL)dropwise to a solution of 2-bromo-4′-methoxyacetonephenone (8.0 g, 34.9mmol) in acetone (100 mL) at room temperature under nitrogen. Stir themixture for 12 hours. Collect the solids by filtration and wash withcold acetone (30 mL) to provide thioacetimidic acid2-(4-methoxyphenyl)-2-oxoethyl ester hydrobromide (10.25 g, 96%).

Heat a mixture of thioacetimidic acid 2-(4-methoxyphenyl)-2-oxoethylester hydrobromide (10.0 g, 32.9 mmol) and zinc (II) chloride (4.50 g,33.0 mmol) in methanol (80 mL) at reflux under nitrogen protection for6.5 hours. Cool the mixture, slowly dilute with saturated NaHCO₃ (300mL), and extract with methylene chloride (400 mL×2). Dry the combinedorganic extracts over sodium sulfate and remove the solvent underreduced pressure. Purify the crude product by flash columnchromatography on silica gel, eluting with hexanes/ethyl acetate (9:1),to provide 4-(4-methoxyphenyl)-2-methylthiazole (6.24 g, 92%): APCI massspectrum m/z 206 [C₁₁H₁₁NOS+H]⁺.

Add a solution of tert-butyllithium (26.35 mmol, 15.5 mL, 1.7 M inhexanes) dropwise to a solution of 4-(4-methoxyphenyl)-2-methylthiazole(6.15 g, 29.9 mmol) in degassed anhydrous tetrahydrofuran (THF) (100 mL)at −78° C. under nitrogen and stir the solution for 45 minutes. To thissolution, add a solution of (6-bromohexyloxy)-tert-butyldimethylsilane(7.20 g, 24.4 mmol) over 5 min and stir the mixture for 2 hours. Warmthe mixture to 0° C., dilute with NH₄Cl (200 mL) and brine (250 mL) andextract with methylene chloride (3×150 mL). Dry the combined organicextracts over magnesium sulfate and remove the solvent under reducedpressure. Purify the crude product by flash column chromatography onsilica gel, eluting with hexanes/ethyl acetate (5:1), to provide2-[7-(tert-butyldimethylsilanyloxy)heptyl]-4-(4-methoxyphenyl)thiazole(6.27 g, 50%).

Add a solution of 1 N tetra-n-butylammonium fluoride (25.0 mmol, 25 mL,1 M solution in THF) dropwise over 10 minutes to a solution of2-[7-(tert-butyldimethylsilanyloxy)heptyl]-4-(4-methoxyphenyl)thiazole(6.27 g, 14.9 mmol) in anhydrous THF (50 mL) at 0° C. under nitrogen andstir the mixture for 30 minutes at 0° C. and then stir at roomtemperature for 3 hours. Dilute the mixture with brine (150 mL) andextract with ethyl acetate (100 mL×3). Dry the combined organic extractsover magnesium sulfate and remove the solvent under reduced pressure.Purify the crude product by flash column chromatography on silica gel,eluting with ethyl acetate/hexanes (1:2), to give7-[4-(4-methoxyphenyl)thiazol-2-yl]heptan-1-ol (4.07 g, 89%): APCI massspectrum m/z 306 [C₁₇H₂₃NO₂S+H]⁺.

Add anhydrous dimethyl sulfoxide (0.25 mL, 3.52 mmol) dropwise over 2minutes to a solution of oxalyl chloride (393 mg, 3.10 mmol) inmethylene chloride (10 mL) at −78° C. under nitrogen and stir themixture for 20 minutes. Add a solution of7-[4-(4-methoxyphenyl)thiazol-2-yl]heptan-1-ol (0.609 g, 1.99 mmol) inmethylene chloride (10 mL) dropwise in 5 minutes and then stir themixture for 30 minutes. To this mixture, add triethylamine (1.0 mL, 7.2mmol), stir and warm the reaction mixture to room temperature for 40minutes. Dilute the mixture with ethyl acetate (100 mL), wash with brine(3×30 mL), dry over sodium sulfate and remove the solvent under reducedpressure to provide 7-[4-(4-methoxyphenyl)thiazol-2-yl]heptanal (0.6 g,99%): APCI mass spectrum m/z 304 [C₁₇H₂₁NO₂S+H]⁺.

Example 31 7-[4-(4-Methoxyphenyl)thiazol-2-yl]heptanoic Acid

Add 2-methyl-2-butene (7.0 mL) and sodium hypochlorite (2.51 g, 27.75mmol) to a solution of 7-[4-(4-methoxyphenyl)thiazol-2-yl]heptanal (4.02g, 13.25 mmol) and potassium dihydrogen phosphate (3.10 g, 22.78 mmol)in tert-butanol (60 mL) and water (12 mL) at room temperature. Stir themixture for 40 minutes, dilute with ethyl acetate (500 mL) and wash withbrine (3×200 mL). Dry the combined organic extracts over sodium sulfateand remove the solvent under reduced pressure. Purify the residue byflash column chromatography on silica gel, eluting withmethanol/methylene chloride (1:19), and triturate the residue withhexanes/methylene chloride to afford7-[4-(4-methoxyphenyl)thiazol-2-yl]heptanoic acid (3.91 g, 88%): APCI MSm/z 320 [C₁₇H₂₁NO₃S+H]⁺.

Prepare Examples 32 and 33, compounds of formula II(b) listed in Table 2below, by the same process as in the preparation of Example 31.

TABLE 2 II(b)

Compounds of formula II(b) Example R¹ n mass spectrum m/z  32 OCH₃ 6 318[C₁₇H₂₁NO₃S − H]⁻  33 OH 6 304 [C₁₆H₁₉NO₃S − H]⁻ 181 OH 4 276[C₁₄H₁₅NO₃S − H]⁻

Preparation 4 7-(2-Oxo-2-phenylethylcarbamoyl)heptanoic Acid MethylEster

Add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) (8.5 g, 44.3 mL)to a solution of 2-amino-1-phenylethanol (5.0 g, 36.4 mmol), subericacid monomethyl ester (6.85 g, 36.4 mmol) and 1-hydroxybenzotriazole(HOBt, 5.0 g, 37.0 mmol) in THF (200 mL) at room temperature undernitrogen. Stir the mixture for 12 hours. Dilute the mixture with ethylacetate (600 mL), wash with 1N HCl (2×150 mL), brine (2×150 mL), NaHCO₃(2×150 mL) and brine (150 mL) solutions and dry over sodium sulfate.Remove the solvent under reduced pressure to provide7-(2-hydroxy-2-phenylethylcarbamoyl)heptanoic acid methyl ester (10.3 g,91%), which is used in the following step without purification.

Add Dess-Martin periodinane (16.5 g, 38.7 mmol) to a solution of7-(2-hydroxy-2-phenylethylcarbamoyl)heptanoic acid methyl ester (10.2 g,33.3 mmol) in methylene chloride (360 mL) at 0° C. under nitrogen, stirand warm the mixture to room temperature for 4 hours. Filter the mixturethrough Celite, wash with ethyl acetate (3×100 mL) and remove thesolvent under reduced pressure. Purify the residue by flash columnchromatography on silica gel, eluting with hexane/ethyl acetate (60:40),to provide 7-(2-oxo-2-phenylethylcarbamoyl)heptanoic acid methyl ester(7.33 g, 72%).

Example 34 7-(5-Phenyloxazol-2-yl)heptanoic Acid Methyl Ester

Add a solution of 7-(2-oxo-2-phenylethylcarbamoyl)heptanoic acid methylester (7.05 g, 23.1 mmol) and carbon tetrabromide (11.3 g, 34.3 mmol) inmethylene chloride over 40 minutes to a mixture of triphenylphosphine(9.0 g, 34.3 mmol) and DMAP (5.51 g, 45.1 mmol) in methylene chloride(500 mL) at room temperature under nitrogen. Stir the mixture for 30minutes and add additional triphenylphosphine (2.6 g, 9.92 mmol) andcarbon tetrabromide (3.35 g, 10.1 mmol). Stir the mixture for anadditional 20 minutes, filter through Celite and wash with ethyl acetate(3×100 mL). Remove the solvent under reduced pressure and purify theresidue by flash column chromatography on silica gel, eluting withhexanes/ethyl acetate (70:30), to provide7-(5-phenyloxazol-2-yl)heptanoic acid methyl ester (2.75 g, 41%).

Example 35 7-(5-Phenyloxazol-2-yl)heptanoic Acid

Add a solution of sodium hydroxide (1.80 g, 45.0 mmol) in water (30 mL)to a solution of 7-(5-phenyloxazol-2-yl)heptanoic acid methyl ester (9.0g, 31.3 mmol) in methanol (30 mL) at room temperature and stir themixture for 4 hours. Adjust the pH of the mixture to 2 with 1 N HCl andextract with ethyl acetate (500 mL). Wash the combined organic layerswith water (3×100 mL), dry over sodium sulfate and remove the solventunder reduced pressure. Crystallize the residue from ethylacetate/hexanes to afford 7-(5-phenyloxazol-2-yl)heptanoic acid (7.3 g,85%): APCI mass spectrum m/z 272 [C₁₆H₁₉NO₃—H]⁻.

Prepare Examples 36-41, compounds of formula II(c) listed in Table 3below, by the same process as in the preparation of Example 35.

TABLE 3 II(c)

Compounds of formula II(c) R¹ (position on R² (position on Example ring)ring) n mass spectrum m/z 36 OCH₃ (1) H 6 302 [C₁₇H₂₁NO₄ − H]⁻ 37 OH (1)Cl (4) 6 321 [C₁₆H₁₈ClNO₄ − H]⁻ 38 OCH₃ (1) OCH₃ (4) 6 332 [C₁₈H₂₃NO₅ −H]⁻ 39 OH (1) H 4 260 [C₁₄H₁₅NO₄ − H]⁻ 40 OCH₃ (1) H 6 302 [C₁₇H₂₁NO₄ −H]⁻ 41 OH (1) H 6 288 [C₁₆H₁₉NO₄ − H]⁻

7-[2-(5-Chloro-2-isopropoxyphenyl)-2-oxoethylcarbamoyl]heptanoic AcidMethyl Ester

Add 2-iodopropane (63.3 mL, 633 mmol) dropwise to a suspension of1-(5-chloro-2-hydroxyphenyl)ethanone (90.0 g, 528 mmol) and potassiumcarbonate (109.46 g, 792 mmol) in DMF (1000 mL) at room temperatureunder nitrogen and heat the mixture at 80° C. for 22 hours. Cool andfilter the mixture and remove the solvent reduced pressure. Dilute theresidue with ethyl acetate (1 L), wash with water (300 mL) and brine(200 mL), dry over sodium sulfate, and remove the solvent under reducedpressure to afford 1-(5-chloro-2-isopropoxyphenyl)ethanone (104.78 g,93%).

Add Copper(II) bromide (199 g, 891 mmol) portionwise to a solution of1-(5-chloro-2-isopropoxyphenyl)ethanone (94.74 g, 446 mmol) in ethylacetate (500 mL) and chloroform (500 mL) at room temperature undernitrogen. Heat the mixture at reflux for 4.5 hours. Cool the mixture andvacuum filter through a plug of Celite, washing with ethyl acetate (1L). Remove the solvents under reduced pressure to provide2-bromo-1-(5-chloro-2-isopropoxyphenyl)ethanone (128.46 g, 98%).

Add hexamethylenetetramine (36.14 g, 258 mmol) to a solution of2-bromo-1-(5-chloro-2-isopropoxyphenyl)ethanone (75.17 g, 258 mmol) inchloroform (400 mL) at room temperature under nitrogen and stir for 2days. Collect the solids by filtration, wash with diethyl ether, and dryovernight under reduced pressure. Suspend the solids in methanol (350ml), cool to 0° C., and treat slowly with concentrated HCl (113 ml, 1365mmol). Warm the mixture to room temperature and stir for 40 hours. Thenheat the mixture to 55° C. for an additional 4 hours. Remove the solidsby filtration, and remove the filtrate solvent under reduced pressure toprovide a solid. Triturate the solid with diethyl ether. Collect theresulting material by filtration to provide2-amino-1-(5-chloro-2-isopropoxyphenyl)ethanone hydrochloride, which isused in the next step without purification.

Add diisopropylethylamine (99 ml, 568 mmol) dropwise to a solution ofEDC HCl (38.01 g, 198 mmol), HOBt (19.19 g, 142 mmol) and octanedioicacid monomethyl ester (53.44 g, 1.42 mmol) in methylene chloride (800mL) at 0° C. under nitrogen. Warm the mixture to room temperature andstir for 1 hour. Add 2-amino-1-(5-chloro-2-isopropoxyphenyl)ethanonehydrochloride (53.44 g, 142 mmol) to the mixture and stir for 18 hours.Remove the solvent under reduced pressure, dilute the residue in ethylacetate (300 mL), wash with water (100 mL) and brine (100 mL), and dryover magnesium sulfate. Remove the solvent under reduced pressure andpurify the residue by flash column chromatography on silica gel, elutingwith hexanes/ethyl acetate (4:6 to 0:10), to afford7-[2-(5-chloro-2-isopropoxyphenyl)-2-oxoethylcarbamoyl]-heptanoic acidmethyl ester (23.87 g, 23% over three steps).

Example 42 7-[5-(5-Chloro-2-isopropoxyphenyl)thiazol-2-yl]heptanoic AcidMethyl Ester

Add Lawesson's reagent (31.03 g, 77 mmol) to a solution of7-[2-(5-chloro-2-isopropoxyphenyl)-2-oxoethylcarbamoyl]heptanoic acidmethyl ester (21.80 g, 55 mmol) in THF (550 mL) at room temperatureunder nitrogen. Heat the mixture at reflux for 3 hours. Remove thesolvent under reduced pressure and purify the residue by flash columnchromatography on silica gel, eluting with ethyl acetate/hexanes (1:3),to afford 7-[5-(5-chloro-2-isopropoxyphenyl)thiazol-2-yl]heptanoic acidmethyl ester (9.96 g, 46%).

Example 43 7-[5-(5-Chloro-2-hydroxyphenyl)thiazol-2-yl]heptanoic AcidMethyl Ester

Add aluminum(III) chloride (6.67 g, 50 mmol) portionwise to a solutionof 7-[5-(5-chloro-2-isopropoxyphenyl)thiazol-2-yl]heptanoic acid methylester (9.90 g, 25 mmol) in methylene chloride (300 mL) at 0° C. undernitrogen. Slowly warm the mixture to room temperature and stir for anadditional 30 minutes. Cool the mixture to 0° C., treat with saturatedaqueous sodium sulfate Na₂SO₄ (150 ml), and stir for 1 hour. Remove thesolvent under reduced pressure, dilute the residue with ethyl acetate(300 mL), wash with water (100 mL) and brine (100 mL), and dry oversodium sulfate. Remove the solvent under reduced pressure and purify theresidue by flash column chromatography on silica gel, eluting with ethylacetate/hexanes (1:9 to 1:1), to afford7-[5-(5-chloro-2-hydroxyphenyl)thiazol-2-yl]heptanoic acid methyl ester(5.67 g, 64%).

Example 44 7-[5-(5-Chloro-2-hydroxy-phenyl)-thiazol-2-yl]-heptanoic acid

Add a solution of sodium hydroxide (2.60 g, 65 mmol) in water (50 mL) toa solution of 7-[5-(5-chloro-2-hydroxyphenyl)thiazol-2-yl]heptanoic acidmethyl ester (5.76 g, 16 mmol) in methanol (100 mL) at 0° C. undernitrogen, warm the mixture to room temperature, and stir for a total of1.5 hours. Remove the solvent under reduced pressure, dilute the residuewith water (200 mL), cool to 0° C., and acidify to pH 1 with 1 N HCl.Collect the precipitate by filtration to afford7-[5-(5-chloro-2-hydroxyphenyl)thiazol-2-yl]heptanoic acid (5.21 g,95%): APCI mass spectrum m/z 338 [C₁₆H₁₈ClNO₃S—H]⁻.

Prepare Examples 45-47, compounds of formula II(d) listed in Table 4below, by the same process as in the preparation of Example 44.

TABLE 4 II(d)

Compounds of formula II(d) Example R¹ n mass spectrum m/z 45 OCH₃ 6 320[C₁₇H₂₁NO₃S + H]⁺ 46 OH 6 304 [C₁₆H₁₉NO₃S − H]⁻ 47 OH 4 276 [C₁₄H₁₅NO₃S− H]⁻

Preparation 6 7-[2-(2-Methoxyphenyl)-2-oxoethylcarbamoyl]heptanoic AcidMethyl Ester

Add triethylamine (8.1 g, 79.9 mmol) dropwise to a solution of2-amino-1-(2-methoxyphenyl)ethanone hydrochloride (13.6 g, 67.4 mmol)and octanedioic acid monomethyl ester (14.0 g, 74.2 mmol) in methylenechloride (600 mL) at 0° C. under nitrogen, and then add EDC HCl (15.5 g,81.0 mmol). Stir the mixture for 4 hours and warm to room temperaturewith stirring for an additional 18 hours. Dilute the mixture in ethylacetate (1.2 L), wash sequentially with water (300 mL), 1 N HCl (300mL), brine (300 mL), saturated sodium, bicarbonate solution (300 mL) andbrine (300 mL), and dry over sodium sulfate. Remove the solvent underreduced pressure to afford7-[2-(2-methoxyphenyl)-2-oxoethylcarbamoyl]heptanoic acid methyl ester(20.0 g, 88%), which is used in the next step without furtherpurification.

Example 48 7-[5-(2-Methoxyphenyl)-1H-imidazol-2-yl]heptanoic Acid MethylEster

Heat a mixture of ammonium acetate (16.5 g, 214 mmol) and7-[2-(2-methoxyphenyl)-2-oxoethylcarbamoyl]heptanoic acid methyl ester(14.2 g, 42.3 mmol) in acetic acid (300 mL) at reflux under nitrogen for15 hours. Remove the solvent under reduced pressure. Dilute the residuein ethyl acetate (500 mL) and adjust to pH 8 with saturated aqueoussodium bicarbonate solution. Extract the aqueous layer with additionalethyl acetate (200 mL) and dry the combined organic extracts over sodiumsulfate and remove the solvent under reduced pressure. Purify theresidue by flash column chromatography on silica gel, eluting with ethylacetate, to afford 7-[5-(2-methoxyphenyl)-1H-imidazol-2-yl]heptanoicacid methyl ester (5.86 g, 44%): APCI mass spectrum m/z 317[C₁₈H₂₄N₂O₃+H]⁺.

Example 49 7-[5-(2-Methoxyphenyl)-1H-imidazol-2-yl]heptanoic Acid

Add a solution of sodium hydroxide (1.85 g, 46 mmol) in water (40 mL) toa solution of 7-[5-(2-methoxyphenyl)-1H-imidazol-2-yl]heptanoic acidmethyl ester (5.84 g, 18.5 mmol) in methanol (30 mL) at room temperatureunder nitrogen and heat the mixture at 40° C. for 4.5 hours. Cool themixture and treat with 1 N HCl (46 mL) and heat at reflux for 30minutes. Collect the precipitate, wash with water (3×30 mL), and dryunder reduced pressure for 12 hours. Triturate the solid with methylenechloride (50 mL) at reflux for 40 min and collect by filtration toprovide 7-[5-(2-methoxyphenyl)-1H-imidazol-2-yl]heptanoic acid (4.27 g,77%). APCI mass spectrum m/z 301 [C₁₇H₂₂N₂O₃—H]⁻.

Prepare Examples 50-55, compounds of formula II(e) listed in Table 5below, by the same process as in the preparation of Example 49.

TABLE 5 II(e)

Compounds of formula II(e) R¹ (position on Example ring) n mass spectrumm/z 50 H 6 271 [C₁₆H₂₀N₂O₂ − H]⁻ 51 OCH₃ (3) 6 301 [C₁₇H₂₂N₂O₃ − H]⁻ 52OH (3) 6 287 [C₁₆H₂₀N₂O₃ − H]⁻ 53 OH (1) 6 287 [C₁₆H₂₀N₂O₃ − H]⁻ 54 OCH₃(1) 4 273 [C₁₅H₁₈N₂O₃ − H]⁻ 55 OH (1) 4 282 [C₁₄H₁₅N₂NaO₃ − H]⁻

Preparation 7 Thiobenzoic Acid Hydrazide

Add a solution of thiobenzoylsulfanylacetic acid (5.5 g, 26.0 mmol) inmethanol (100 mL) and to a solution of thionyl chloride (52 mL) at roomtemperature under nitrogen and heat the mixture at reflux for 12 hours.Remove the solvent under reduced pressure, dissolve the residue in ethylacetate (200 mL), wash with saturated NaHCO₃ (200 mL) and brine (200 mL)solutions, and dry over sodium sulfate. Remove the solvent under reducedpressure to provide thiobenzoylsulfanylacetic acid methyl ester (5.7 g,97%).

Add a solution of thiobenzoylsulfanylacetic acid methyl ester (1.9 g,8.4 mmol) in ethanol (30 mL) to a solution of anhydrous hydrazine (1 mL)at room temperature under nitrogen and stir for 2 hours. Then add water(20 mL) and remove the solvent under reduced pressure. Dissolve theresidue in ethyl acetate (300 mL), wash with water (200 mL) and brine(200 mL), and dry over magnesium sulfate. Remove the solvent underreduced pressure to provide thiobenzoic acid hydrazide (1.2 g, 94%).

Preparation 8 7-Ethoxycarbonimidoylheptanoic Acid Ethyl EsterHydrochloride

Add sodium cyanide (12.5 g, 255 mmol) and tetra-n-butylammonium iodide(10 g, 27.0 mmol) portionwise to a solution of 7-bromoheptanoic acidmethyl ester (25 g, 105 mmol) in DMSO (300 mL) at room temperature undernitrogen and heat the mixture at 50° C. for 4 hours. Cool the mixtureand dilute with water (200 mL) and extract with diethyl ether (2×200mL). Dry the combined organic extracts over sodium sulfate and removethe solvent under reduced pressure to provide 7-cyanoheptanoic acidethyl ester (18.2 g, 94%).

Bubble hydrogen chloride gas into a solution of 7-cyanoheptanoic acidethyl ester (3.7 g, 20.0 mmol) in ethanol (24 mL, 40 mmol) and diethylether (100 mL) at 0° C. for 15 minutes. Remove the solvent under reducedpressure to provide 7-ethoxycarbonimidoylheptanoic acid ethyl ester (5.4g, >99%), which is used without further purification.

Example 56 7-(5-Phenyl[1,3,4]thiadiazol)heptanoic Acid Ethyl Ester

Heat a solution of thiobenzoic hydrazide (1.2 g, 7.90 mmol) and7-ethoxycarbonimidoylheptanoic acid ethyl ester (2.9 g, 11.0 mmol) inethanol (35 mL) at reflux under nitrogen for 3 hours. Remove the solventunder reduced pressure. Dissolve the residue in ethyl acetate (200 mL),wash with water (200 mL) and brine (200 mL), and dry over sodiumsulfate. Remove the solvent under reduced pressure and purify theresidue by flash column chromatography on silica gel, eluting withhexane/ethyl acetate (4:1), to provide7-(5-phenyl[1,3,4]thiadiazol)heptanoic acid ethyl ester (1.15 g, 45%).

Example 57 7-(5-Phenyl[1,3,4]thiadiazol-2-yl)heptanoic Acid

Add a solution of potassium hydroxide (1.2 g 23 mmol) in water (50 mL)to a solution of 7-(5-phenyl-[1,3,4]thiadiazol)heptanoic acid ethylester (3.4 g, 11 mmol) in THF (30 mL) and methanol (30 mL) at roomtemperature under nitrogen and heat the mixture at reflux for 3 hours.Remove the solvent under reduced pressure, dilute the residue with water(200 mL) and wash with ethyl acetate (200 mL). Adjust the pH of theaqueous layer to 3 with concentrated HCl and extract with ethyl acetate(3×200 mL). Wash the combined organic extracts with brine (200 mL), dryover sodium sulfate and remove the solvent under reduced pressure toafford 7-(5-phenyl[1,3,4]thiadiazol-2-yl)heptanoic acid (2.9 g, 93%).APCI mass spectrum m/z 289 [C₁₅H₁₈N₂O₂S—H]⁻.

Prepare Examples 58-61, compounds of formula II(f) listed in Table 6below, by the same process as in the preparation of Example 57.

TABLE 6 II(f)

Compounds of formula II(f) Example R² (position on ring) n mass spectrumm/z 58 OCH₃ (3) 6 335 [C₁₆H₂₀N₂O₄S − H]⁻ 59 H 6 305 [C₁₅H₁₈N₂O₃S − H]⁻60 Cl (4) 6 341 [C₁₅H₁₇ClN₂O₃S + H]⁺ 61 H 4 277 [C₁₃H₁₄N₂O₃S − H]⁻

Preparation 9 8-Amino-8-(benzoylhydrazono)octanoic Acid Ethyl Ester

Add triethylamine (5.6 mL, 40 mmol) to a solution of7-ethoxycarbonimidoylheptanoic acid ethyl ester (11.0 g, 41 mmol) andbenzoic acid hydrazide (5.5 g, 40 mmol) in ethanol (110 mL) at roomtemperature under nitrogen and stir the mixture for 12 hours. Remove thesolvent under reduced pressure, dissolve the residue in ethyl acetate(200 mL), wash with saturated NaHCO₃ (200 mL) and brine (200 mL)solutions, and dry over sodium sulfate. Remove the solvent under reducedpressure to provide 8-amino-8-(benzoylhydrazono)octanoic acid ethylester (8.3 g, 64%).

Example 62 7-(5-Phenyl-4H-[1,3,4]triazol-3-yl)heptanoic Acid Ethyl Ester

Heat a solution of 8-amino-8-(benzoylhydrazono)octanoic acid ethyl ester(4.2 g, 26 mmol) in o-xylene (400 mL) at reflux under nitrogen for 5hours and then remove the solvent under reduced pressure. Dilute theresidue with ethyl acetate (500 mL), wash with saturated NaHCO₃ (200 mL)and brine (200 mL) solutions and dry over sodium sulfate. Remove thesolvent under reduced pressure and purify the residue by flash columnchromatography on silica gel, eluting with methanol/methylene chloride(1:9), to provide 7-(5-phenyl-4H-[1,3,4]triazol-3-yl)heptanoic acidethyl ester (2.2 g, 58%).

Example 63 7-(5-Phenyl-4H-[1,3,4]triazol-3-yl)-heptanoic acid

Add a solution of potassium hydroxide (1.8 g, 32 mmol) in water (70 mL)to a solution of 7-(5-phenyl-4H-[1,3,4]triazol-3-yl)heptanoic acid ethylester (4.9 g, 16 mmol) in THF (50 mL) and methanol (50 mL) at roomtemperature under nitrogen and heat the mixture at reflux for 3 hours.Remove the solvent under reduced pressure, dilute the residue with water(200 mL) and wash with ethyl acetate (200 mL). Adjust the pH of theaqueous layer to 3 with concentrated HCl and extract with ethyl acetate(3×200 mL). Wash the combined organic extracts with brine (200 mL), dryover sodium sulfate and remove the solvent under reduced pressure toafford 7-(5-phenyl-4H-[1,3,4]triazol-3-yl)heptanoic acid (4.4 g, 99%).APCI mass spectrum m/z 273 [C₁₅H₁₉N₃O₂]⁻.

Prepare Examples 64-39, compounds of formula II(g) listed in Table 7below, by the same process as in the preparation of Example 63.

TABLE 7 II(g)

Compounds of formula II(g) Example R² (position on ring) R⁸ n massspectrum m/z 64 OCH₃ (3) H 6 319 [C₁₆H₂₁N₃O₄ − H]⁻ 65 H H 6 289[C₁₅H₁₉N₃O₃ − H]⁻ 66 Cl (4) H 6 323 [C₁₅H₁₈ClN₃O₃ − H]⁻ 67 H H 4 261[C₁₃H₁₅N₃O₃]⁻ 68 H CH₃ 4 277 [C₁₄H₁₇N₃O₃]⁻

Preparation 10 N-Hydroxy-2-methoxybenzamidine

Add potassium hydroxide (30.3 g, 225 mmol) to a solution of2-methoxybenzonitrile (25.0 g, 187 mmol) and hydroxylamine hydrochloride(15.77 g, 225 mmol) in ethanol (500 mL) at room temperature undernitrogen and heat the mixture at reflux for 12 hours. Remove the solventunder reduced pressure, triturate the residue with ethyl acetate/hexanes(1:9, 300 mL) and collect by vacuum filtration to provideN-hydroxy-2-methoxybenzamidine (24.0 g, 91%).

Example 69 5-[3-(2-Methoxyphenyl)-[1,2,4]oxadiazol-5-yl]pentanoic AcidMethyl Ester

Add 5-chlorocarbonylpentanoic acid methyl ester (15.30 g, 86 mmol) to asolution of N-hydroxy-2-methoxybenzamidine (12.0 g, 71 mmol) in pyridine(40 mL) and under nitrogen at a rate to keep the mixture at a gentlereflux. Then, heat the mixture at reflux for 4 hours. Dilute the mixturewith water (300 mL) and extract with methylene chloride (3×200 mL). Washthe combined organic extracts with brine (100 mL), dry over sodiumsulfate and remove the solvent under reduced pressure. Purify theresidue by flash column chromatography on silica gel, eluting with ethylacetate/hexanes (1:19), to afford5-[3-(2-methoxyphenyl)-[1,2,4]oxadiazol-5-yl]pentanoic acid methyl ester(12.8 g, 55%).

Example 70 5-[3-(2-Methoxyphenyl)-[1,2,4]oxadiazol-5-yl]pentanoic Acid

Add 2 N sodium hydroxide (20 mL) to a solution of5-[3-(2-methoxyphenyl)-[1,2,4]oxadiazol-5-yl]pentanoic acid methyl ester(4.00 g, 13 mmol) in methanol (100 mL) at room temperature undernitrogen and stir the mixture for 3 hours. Remove the solvent underreduced pressure, dilute the residue with water (200 mL) and wash withdiethyl ether (200 mL). Adjust the aqueous layer to pH 1 with 2 N HCland collect the solids by vacuum filtration to afford5-[3-(2-methoxyphenyl)-[1,2,4]oxadiazol-5-yl]pentanoic acid (3.65 g,99%). APCI mass spectrum m/z 275 [C₁₄H₁₆N₂O₄—H]⁻.

Prepare Examples 71-73, compounds of formula II(h) listed in Table 8below, by the same process as in the preparation of Example 40.

TABLE 8 II(h)

Compounds of formula II(h) R¹ (position on Example ring) N mass spectrumm/z 71 OH (1) 4 261 [C₁₃H₁₄N₂O₄ − H]− 72 CH₃ (2) 7 301 [C₁₇H₂₂N₂O₃ − H]−73 CF₃ (3) 7 355 [C₁₇H₁₉F₃N₂O₃ − H]−

Preparation 11 4-(2-Isopropoxy-phenyl)-1H-imidazole

Add tetrakis(triphenylphosphine)palladium(0) (500 mg) to a degassedsuspension of 4-bromo-1H-imidazole (5.0 g, 34 mmol) and2-isopropoxyphenyl boronic acid (9.19 g, 51 mmol) in dioxane (250 mL)and 2 M sodium carbonate solution (10.81 g, 102 mmol) at roomtemperature under nitrogen and heat the mixture at reflux for 21 hours.Remove the solvent under reduced pressure, dilute the residue with ethylacetate (500 mL) and filter through a plug of Celite. Dry the filtrateover sodium sulfate, treat with silica gel (20 g) and remove the solventunder reduced pressure. Purify the residue by flash columnchromatography on silica gel, eluting with ethyl acetate, to affordcrude 4-(2-isopropoxyphenyl)-1H-imidazole (5.01 g, 73%) which is usedwithout further purification in the next step.

Example 74 8-[4-(2-Isopropoxyphenyl)imidazol-1-yl]octanoic Acid MethylEster

Add sodium hydride (1.82 g, 38 mmol) to a suspension of4-(2-isopropoxyphenyl)-1H-imidazole (5.01 g, 25 mmol) in THF (125 mL) at0° C. under nitrogen, and warm the mixture to room temperature and stirfor 1 hour. Cool the mixture to 0° C. and add 8-bromooctanoic acidmethyl ester (5.98 g, 25 mmol) and tetra-n-butylammonium iodide (0.55 g,1.5 mmol) and warm the mixture to room temperature to stir for 8 hours.Dilute the mixture with water (20 mL) and remove the solvent underreduced pressure. Dilute the residue with ethyl acetate (300 mL), washwith water (100 mL) and brine (100 mL), dry over sodium sulfate, andremove the solvent under reduced pressure to provide8-[4-(2-isopropoxyphenyl)imidazol-1-yl]octanoic acid methyl ester (5.16g, 57%), which is used in the next step without further purification.

Example 75 8-[4-(2-Hydroxyphenyl)imidazol-1-yl]octanoic Acid MethylEster

Add aluminum(III) chloride (3.84 g, 29 mmol) to a suspension of8-[4-(2-isopropoxyphenyl)imidazol-1-yl]octanoic acid methyl ester (5.16g, 14 mmol) in methylene chloride (150 mL) at 0° C. under nitrogen. Warmthe mixture to room temperature and stir for 6 hours. Dilute the mixturewith saturated aqueous sodium sulfate (50 mL) and remove the solventunder reduced pressure. Dilute the residue with ethyl acetate (300 mL),wash with brine (100 mL), dry over sodium sulfate, and remove thesolvent under reduced pressure. Purify the residue by flash columnchromatography on silica gel, eluting with ethyl acetate/hexanes (2:8 to3:7), to provide 8-[4-(2-hydroxyphenyl)imidazol-1-yl]octanoic acidmethyl ester (2.63 g, 57%).

Example 76 8-[4-(2-Hydroxyphenyl)imidazol-1-yl]octanoic Acid

Add sodium hydroxide (1.32 g, 33 mmol) in water (20 mL) to a suspensionof 8-[4-(2-hydroxyphenyl)imidazol-1-yl]octanoic acid methyl ester (2.60g, 8 mmol) in methanol (50 mL) at 0° C. under nitrogen and warm themixture to room temperature and stir for 8 hours. Remove the solventunder reduced pressure, dilute the residue with water (200 mL), cool to0° C., and acidify to pH 1 with 1 N HCl. Collect the precipitate toprovide 8-[4-(2-hydroxyphenyl)imidazol-1-yl]octanoic acid (1.70 g, 68%).APCI mass spectrum m/z 301 [C₁₇H₂₂N₂O₃—H]⁻.

Prepare Examples 77 and 78, compounds of formula II(i) listed in Table 9below, by the same process as in the preparation of Example 76.

TABLE 9 II(i)

Compounds of formula II(i) Example R¹ n mass spectrum m/z 77 H 7 285[C₁₇H₂₂N₂O₂ − H]⁻ 78 OH 4 259 [C₁₄H₁₆N₂O₃ − H]⁻

Example 79 7-[3-(2-Hydroxyphenyl)pyrazol-1-yl]heptanoic Acid Ethyl Ester

Add sodium hydride (1.50 g, 31 mmol, 60% suspension in mineral oil) to asuspension of 2-(1H-pyrazol-3-yl)phenol (5.0 g, 31 mmol) and7-bromoheptanoic acid ethyl ester (7.4 g, 31 mmol) in DMF (75 mL) atroom temperature under nitrogen and heat the mixture at 75° C. for 16hours. Remove the solvent under reduced pressure, dilute the residuewith ethyl acetate (300 mL), wash with water (100 mL), and dry oversodium sulfate. Remove the solvent under reduced pressure and purify theresidue by flash column chromatography on silica gel, eluting withhexanes/ethyl acetate (9:1), to provide7-[3-(2-hydroxyphenyl)pyrazol-1-yl]heptanoic acid ethyl ester (4.73 g,48%).

Example 80 7-[3-(2-Methoxyphenyl)pyrazol-1-yl]heptanoic Acid Ethyl Ester

Add sodium hydride (900 mg, 18 mmol, 60% suspension in mineral oil) to asuspension of 7-[3-(2-hydroxyphenyl)pyrazol-1-yl]heptanoic acid ethylester (4.73 g, 15 mmol) and iodomethane (1.1 mL, 18 mmol) in THF (70 mL)at 0° C. under nitrogen and warm the mixture to room temperature to stirfor 12 hours. Remove the solvent under reduced pressure, dilute theresidue with ethyl acetate (150 mL), wash with water (100 mL), and dryover sodium sulfate. Remove the solvent under reduced pressure andpurify the residue by flash column chromatography on silica gel, elutingwith hexanes/ethyl acetate (9:1), to provide7-[3-(2-Methoxyphenyl)pyrazol-1-yl]heptanoic acid ethyl ester (3.75 g,76%).

Example 81 7-[3-(2-Methoxyphenyl)pyrazol-1-yl]heptanoic Acid

Add 2 N sodium hydroxide (20 mL) to a solution of7-[3-(2-Methoxyphenyl)pyrazol-1-yl]heptanoic acid ethyl ester (3.75 g,11.4 mmol) in methanol (40 mL) at room temperature under nitrogen andstir the mixture for 8 hours. Remove the solvent under reduced pressure,dilute the residue with water (100 mL), acidify to pH 3 with 1 N HCl,extract with ethyl acetate (200 mL), and dry over sodium sulfate. Removethe solvent under reduced pressure to provide7-[3-(2-methoxyphenyl)pyrazol-1-yl]heptanoic acid (3.06 g, 89%). APCImass spectrum m/z 303 [C₁₇H₂₂N₂O₃+H]⁺.

Prepare Examples 82-87, compounds of formula II(j) listed in Table 10below, by the same process as in the preparation of Example 81.

TABLE 10 II(j)

Compounds of formula II(j) Example R¹ R² (position on ring) n massspectrum m/z 82 OH H 6 289 [C₁₆H₂₀N₂O₃ + H]⁺ 83 OH Cl (4) 6 321[C₁₆H₁₉ClN₂O₃ − H]⁻ 84 OH Br (4) 6 366 [C₁₆H₁₉BrN₂O₃ − H]⁻ 85 OCH3 H 4273 [C₁₅H₁₈N₂O₃ − H]⁻ 86 OH H 4 259 [C₁₄H₁₆N₂O₃ − H]⁻ 87 OH OCH3 (3) 4291 [C₁₅H₁₈N₂O₄ + H]⁺

Example 88 8-(2-Phenylimidazol-1-yl)octanoic Acid Methyl Ester

Add sodium hydride (1.3 g, 42 mmol) to a mixture of2-phenyl-1H-imidazole (5.0 g, 35 mmol), 8-bromooctanoic acid methylester (8.22 g, 35 mmol), potassium carbonate (5.75 g, 42 mmol), andtetra-n-butylammonium iodide (0.77 g, 2 mmol) in DMF (250 mL) at 0° C.under nitrogen. Heat the mixture to 75° C. and stir for 21 hours. Removethe solvent under reduced pressure, dissolve the residue in chloroform(200 mL), wash with water (100 mL) and brine (100 mL), and dry oversodium sulfate. Remove the solvent under reduced pressure and purify theresidue by flash column chromatography on silica gel, eluting withmethanol/methylene chloride (1:9), to afford8-(2-phenylimidazol-1-yl)octanoic acid methyl ester (4.90 g, 47%).

Example 89 8-(2-Phenylimidazol-1-yl)octanoic Acid

Add sodium hydroxide (6.0 g, 150 mmol) in water (50 mL) to a suspensionof 8-(2-phenylimidazol-1-yl)octanoic acid methyl ester (7.60 g, 25 mmol)in methanol (100 mL) at 0° C. under nitrogen. Warm the mixture to roomtemperature and stir for a total of 8 hours. Remove the solvent underreduced pressure, dilute the residue with water (300 mL), cool to 0° C.,and acidify to pH 1 with 1 N HCl. Collect the precipitate and trituratewith hexanes to afford 8-(2-phenylimidazol-1-yl)octanoic acid (4.22 g,52%). APCI mass spectrum m/z 285 [C₁₇H₂₂N₂O₂—H]⁻.

Preparation 12 2-Methoxy-N-hydroxybenzenecarboxyimidoyl Chloride

Add sodium hydroxide (8.50 g, 220 mmol) in water (150 mL) to a solutionof o-anisaldehyde (25.0 g, 180 mmol) and hydroxylamine hydrochloride(15.4 g, 220 mmol) in ethanol (150 mL) and water (150 mL) at roomtemperature and stir the mixture for 3 hours. Acidify the mixture to pH6 with 1 N HCl solution and collect the solids by vacuum filtration toprovide 2-methoxybenzaldehyde oxime (32.0 g, 99%).

Add N-Chlorosuccinimide (8.30 g, 65 mmol) portionwise to a solution of2-methoxybenzaldehyde oxime (10.0 g, 65 mmol) in DMF (100 mL) at roomtemperature under nitrogen. Heat the mixture at 50° C. for 5 hours. Pourthe mixture into ice water (300 mL) collect the solids by vacuumfiltration to provide 2-methoxy-N-hydroxybenzenecarboxyimidoyl chloride(9.80 g, 81%).

Example 90 7-[3-(2-Methoxyphenyl)isoxazol-5-yl]heptanoic Acid MethylEster

Add triethylamine (8.08 g, 80 mmol) to a solution of2-methoxy-N-hydroxybenzenecarboxyimidoyl chloride (8.0 g, 40 mmol) andmethyl 7-oxtynoate (10.50 g, 50 mmol) in THF (100 mL) at roomtemperature and stir the mixture for 24 hours. Dilute the mixture withwater (500 mL) and extract with ethyl acetate (3×200 mL). Wash thecombined organic extracts with water (100 mL) and brine (100 mL) and dryover sodium sulfate. Remove the solvent under reduced pressure andpurify the residue by flash column chromatography on silica gel, elutingwith ethyl acetate/hexanes (1:9), to afford7-[3-(2-methoxyphenyl)isoxazol-5-yl]heptanoic acid methyl ester (7.80 g,55%).

Example 91 7-[3-(2-Methoxyphenyl)isoxazol-5-yl]heptanoic Acid

Add 2 N sodium hydroxide (15 mL) to a solution of7-[3-(2-methoxyphenyl)isoxazol-5-yl]heptanoic acid methyl ester (2.89 g,8 mmol) in methanol (50 mL) at room temperature under nitrogen and stirfor 3 hours. Remove the solvent under reduced pressure, dilute theresidue with water (100 mL), and wash with methyl tert-butyl ether (100mL). Acidify the mixture to pH 1 with 1 N HCl and extract with ethylacetate (3×100 mL). Wash the combined organic extracts with water (100mL) and brine (100), dry over sodium sulfate, and remove the solventunder reduced pressure to provide7-[3-(2-methoxyphenyl)isoxazol-5-yl]heptanoic acid (2.41 g, 98%): APCImass spectrum m/z 302 [C₁₇H₂₁INO₄—H]⁻.

Prepare Examples 92-98, compounds of formula II(k) listed in Table 11below, by the same process as in the preparation of Example 91.

TABLE 11 II(k)

Compounds of formula II(k) R¹ (position on R² (position on Example ring)ring) n mass spectrum m/z 92 OH H 4 260 [C₁₄H₁₅NO₄ − H]⁻ 93 OCH₃ Cl 4309 [C₁₅H₁₆ClNO₄ − H]⁻ 94 OH Cl 4 294 [C₁₄H₁₄ClNO₄ − H]⁻ 95 OH Cl 6 322[C₁₆H₁₈ClNO₄ − H]⁻ 96 OCH₃ Cl 6 337 [C₁₇H₂₀ClNO₄ − H]⁻ 97 OH H 6 288[C₁₆H₁₉NO₄ − H]⁻ 98 OH H 4 260 [C₁₄H₁₅NO₄ − H]⁻

Preparation 13 3-[3-(5-Chloro-2-methoxyphenyl)isoxazol-4-yl]propan-1-ol

Add N-Chlorosuccinimide (8.6 g, 65 mmol) portionwise to a solution ofanisaldehyde oxime (10.0 g, 65 mmol) in DMF (250 mL) at room temperatureunder nitrogen and heat at 50° C. for 6 hours. Pour the mixture into icewater (800 mL) and collect the solids by vacuum filtration to provide5-chloro-2-methoxybenzenecarboxyimidoyl chloride (13.6 g, 92%).

Add triethylamine (10.1 mL, 100 mmol) to a solution of5-chloro-2-methoxybenzenecarboxyimidoyl chloride (10.9 g, 50 mmol) anddihydropyran (4.2 g, 50 mmol) in THF (150 mL) at room temperature andstir the mixture for 48 hours. Dilute the mixture with water (500 mL)and extract with ethyl acetate (3×200 mL). Wash the combined organicextracts with brine (100 mL), dry over sodium sulfate and remove thesolvents under reduced pressure to afford3-(5-chloro-2-methoxyphenyl)-3a,5,6,7a-tetrahydro-4H-pyrano[3,2-d]isoxazole(12.9 g, >99%) that is used in the next step without purification.

Heat a solution of3-(5-chloro-2-methoxyphenyl)-3a,5,6,7a-tetrahydro-4H-pyrano[3,2-d]isoxazole(12.0 g, 44 mmol) in 12 N HCl (200 mL) at 50° C. for 24 hours and thendilute the mixture with water (300 mL) and extract with ethyl acetate(3×200 mL). Wash the combined organic extracts with brine (100 mL), dryover Na₂SO₄ and remove the solvents under reduced pressure. Purify theresidue by flash column chromatography on silica gel, eluting with ethylacetate/hexanes (1:1), to provide3-[3-(5-chloro-2-methoxyphenyl)isoxazol-4-yl]propan-1-ol (8.0 g, 68%).

Example 99 3-[3-(5-Chloro-2-methoxyphenyl)isoxazol-5-yl]propionic Acid

Slowly add sodium perchlorate (2.0 g, 22 mmol) and bleach solution (1mL) to a mixture of3-[3-(5-chloro-2-methoxyphenyl)isoxazol-4-yl]propan-1-ol (3.0 g, 11mmol), 2,2,6,6,-tetramethylpiperidinooxy (50 mg) in acetonitrile (30mL), and saturated potassium phosphate solution (30 mL) at 35° C. andstir the mixture for 12 hours. Adjust the pH of the mixture to pH 8 with2 N NaOH solution and add saturated sodium sulfite solution (40 mL).Wash the mixture with tert-butyl methyl ether (2×20 mL), acidify to pH 1with 1 N HCl and extract with ethyl acetate (3×100 mL). Wash thecombined organic extracts with brine (100 mL), dry over Na₂SO₄ andremove the solvents under reduced pressure to provide3-[3-(5-chloro-2-methoxyphenyl)isoxazol-5-yl]propionic acid (2.65 g,85%): APCI mass, spectrum m/z 280 [C₁₃H₁₂ClNO₄—H]⁻.

Prepare Examples 100-102, compounds of formula II(l) listed in Table 12below, by the same process as in the preparation of Example 99.

TABLE 12 II(l)

Compounds of formula II(l) Example R¹ R² mass spectrum m/z 100 OH H 260[C₁₄H₁₅NO₄ − H]⁻ 101 OCH3 Cl 308 [C₁₅H₁₆ClNO₄ − H]⁻ 102 OH Cl 294[C₁₄H₁₄ClNO₄ − H]⁻

Preparation 14

Add sodium carbonate (4.5 g, 42 mmol) to a solution of commerciallyavailable 5-cyano-pentanoic acid methyl ester (5.0 g, 35 mmol) andhydroxylamine hydrochloride (2.9 g, 42 mmol) in ethanol (100 mL) at roomtemperature under nitrogen and heat the mixture at reflux for 5.5 h.Filter the mixture through celite and remove the solvent under reducedpressure to provide crude methyl 5-(N-hydroxycarbamimidoyl)pentanoate(5.3 g, 87% yield), which is carried forward without furtherpurification.

Example 103 Methyl5-[5-(2-Methoxyphenyl)-[1,2,4]oxadiazol-3-yl]pentanoate

Add 2-methoxybenzoyl chloride (5.20 g, 35 mmol) to a solution of methyl5-(N-hydroxycarbamimidoyl)pentanoate (5.3 g, 34 mmol) in pyridine (50mL) under nitrogen at 0° C. and then heat the mixture at reflux for 6 h.Remove the solvent under reduced pressure, dilute the residue with ethylacetate and wash with 1 N HCl. Wash the combined organic extracts withbrine, dry over sodium sulfate and remove the solvent under reducedpressure. Purify the residue by flash column chromatography on silicagel, eluting with ethyl acetate/hexanes (1:4), to afford methyl5-[5-(2-methoxyphenyl)[1,2,4]oxadiazol-3-yl]pentanoate (3.4 g, 33% overtwo steps): ¹H NMR (CDCl₃) δ 1.7-1.85 (m, 2H), 1.85-2.00 (m, 2H), 2.35(t, 2H), 2.95 (t, 2H), 3.65 (s, 3H), 3.95 (s, 3H), 7.00 (m, 2H), 7.45(t, 1H), 8.00 (d, 1H).

Example 104 5-[5-(2-Methoxyphenyl)[1,2,4]oxadiazol-3-yl]pentanoic Acid

Add a solution of 2 N sodium hydroxide (20 mL) to a solution of methyl5-[5-(2-methoxyphenyl)[1,2,4]oxadiazol-3-yl]pentanoate (3.4 g, 11 mmol)in methanol (50 mL) at room temperature under nitrogen and stir themixture for 3.5 h. Remove the solvent under reduced pressure, dilute theresidue with water (200 mL) and wash with diethyl ether (200 mL). Adjustthe aqueous layer to pH 1 with 1 N HCl and collect the solids by vacuumfiltration to afford5-[5-(2-methoxyphenyl)[1,2,4]oxadiazol-3-yl]pentanoic acid (2.2 g, 72%):APCI MS m/z 275 [C₁₄H₁₆N₂O₄—H]⁻.

Example 105 5-[5-(2-Hydroxyphenyl)[1,2,4]oxadiazol-3-yl]pentanoic Acid

Add boron tribromide (4.9 mL, 51.7 mmol) dropwise to a stirred solutionof methyl 5-[5-(2-methoxyphenyl)[1,2,4]oxadiazol-3-yl]pentanoate (3.0 g,10.3 mmol) in methylene chloride (70 mL) at 0° C. under nitrogen. Allowto warm to room temperature and stir for 5 hours. Cool the mixture to 0°C., add methanol (20 mL) dropwise and allow to warm to room temperature.Remove the solvents under reduced pressure and purify the residue byflash column chromatography on silica gel, eluting with ethylacetate/hexanes (1:9), to provide methyl5-[5-(2-hydroxyphenyl)[1,2,4]oxadiazol-3-yl]pentanoate, which is usedwithout further purification. Add a solution of sodium hydroxide (800mg, 20 mmol) in water (15 mL) to a solution of methyl5-[5-(2-hydroxyphenyl)[1,2,4]oxadiazol-3-yl]pentanoate (2.6 g, 9.4 mmol)in methanol (30 mL) at room temperature and stir the mixture for 2hours. Adjust the pH of the mixture to 2 with 1 N HCl and extract withethyl acetate (2×150 mL). Wash the combined organic extracts with water(3×150 mL), dry over sodium sulfate and remove the solvent under reducedpressure. Triturate the residue with hexanes/ethyl acetate and collectthe solids by vacuum filtration to afford the title compound (2.4 g, 89%over two steps). APCI MS m/z 261 [C₁₃H₁₄N₂O₄—H]⁻.

Preparation 15 5-[2-(2-methoxyphenyl)-2-oxo-ethylcarbamoyl]-pentanoicacid methyl ester

Add sodium azide (2.14 g, 32.92 mmol) to a solution of2-bromo-2′-methoxyacetophenone (5.01 g, 21.87 mmol) in 75 ml of DMSO.Stir the mixture at ambient temperature for 18 hours and dilute it with250 ml of water. Extract the mixture with ether (3×). Dry the combinedorganic layers with MgSO₄. Filter off the drying agent and concentratein vacuo to afford 3.54 g of 2-azido-1-(2-methoxyphenyl)-ethanone.

Dissolve 2-azido-1-(2-methoxy-phenyl)-ethanone (3.54 g, 18.5 mmol) in1328 ml of MeOH and 9 ml of concentrated HCl. Add 943 mg of 10% Pd/C andexpose the reaction mixture to 60 psi of H₂ for 5 hours at ambienttemperature. Filter the catalyst off through a pad of celite andconcentrate the filtrate in vacuo to afford 3.74 g of crude2-amino-1-(2-methoxyphenyl)-ethanone as the hydrochloride salt.

Dissolve 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride(3.57 g, 18.62 mmol), 4-(dimethylamino)pryidine (463.8 mg, 3.79 mmol)and adipic acid monomethyl ester (2.97 g, 18.55 mmol) in 100 mL ofCH₂Cl₂ and allow it to stir at room temperature for 45 minutes. Addcrude 2-amino-1-(2-methoxy-phenyl)-ethanone hydrochloride (3.74 g, 18.55mmol) and triethylamine (3.75 g, 37.1 mmol) to the reaction mixture andallow it to stir at ambient temperature for 22 hours. Dilute thereaction with 200 mL of CH₂Cl₂ and wash it with 1 N HCl (2×), saturatedaqueous NaHCO₃ (2×) and brine (1×). Dry the organic layer with MgSO₄.Filter the drying agent and concentrate in vacuo to afford 4.57 g of thetitled product (80%): mass spectrum: m/z=308.1 (M+H).

Example 106 5-[5-(2-methoxyphenyl)-oxazol-2-yl)-pentanoic acid methylester

Dissolve 5-[2-(2-methoxy-phenyl)-2-oxo-ethylcarbamoyl]-pentanoic acidmethyl ester (4.57 g, 14.87 mmol) and 4-(dimethylamino)pryidine (3.56 g,29.14 mmol) in 150 mL CH₂Cl₂ and cool the reaction mixture in an icebath. Add triphenylphosphinedibromide (12.32 g, 1.96 mmol) to thereaction portionwise over 15 minutes. Raise the reaction to ambienttemperature and allow it to stir for 12 hours.

Wash the reaction with water and brine (2×). Dry the organic fayers withMgSO₄. Filter off the drying agent and concentrate in vacuo to afford acrude residue. Purify the residue using silica gel chromatographyeluting with hexanes/ethyl acetate mixtures to afford 2.47 g of thetitled product (57%): mass spectrum: m/z=290.1 (M+H).

Example 107 5-[5-(2-methoxy-phenyl)-oxazol-2-yl]-pentanoic acid

Dissolve 5-[5-(2-methoxy-phenyl)-oxazol-2-yl]-pentanoic acid methylester (2.47 g, 8.54 mmol) in 40 mL of dioxane and add a solution of LiOH(1.0344 g, 43.19 mmol) in 20 mL of water to the solution. Allow thereaction to stir at ambient temperature for 21 hours. Acidify thereaction with 5N HCl and concentrate the reaction mixture in vacuo toremove the dioxane. Extract the aqueous residue with EtOAc (3×). Dry thecombined organic layers with MgSO₄. Filter off the drying agent andconcentrate in vacuo to afford a crude residue. Recrystallize theresidue in EtOAc/hexanes to afford 1.7692 g of the titled product (75%):mass spectrum: m/z=276.1 (M+H).

Preparation 165-[2-(5-Chloro-2-methoxy-phenyl)-2-oxo-ethylcarbamoyl]-pentanoic acidmethyl ester

Dissolve 5′-chloro-2′-hydroxyacetophenone (15.3594 g, 90.03 mmol) in 100mL of anhydrous acetonitrile in a pressure vessel. Add K₂CO₃ (13.7541 g,99.52 mmol) and MeI (25.56 mL, 180.06 mmol) to the vessel. Seal thevessel and heat the reaction mixture to 85° C. for 18 hours. Cool thereaction and concentrate it in vacuo. Partition the residue between Et₂Oand H₂O. Wash the organic layers with 2N NaOH (2×). Dry the combinedorganic layers with MgSO₄. Filter off the drying agent and concentratein vacuo to afford 13.15 g of 1-(5-chloro-2-methoxy-phenyl)-ethanone(79%).

Suspend CuBr₂ (29.44 g, 131.81 mmol) in 150 mL EtOAc and heat it toreflux. Dissolve 1-(5-chloro-2-methoxy-phenyl)-ethanone (13.15 g, 71.23mmol) in 100 mL of CHCl₃ and add it dropwise to the reaction. After 4hours, filter the reaction to remove the solids. Concentrate thefiltrate in vacuo and dissolve the residue in EtOAc. Wash the organiclayer with saturated aqueous NaHCO₃ and water. Dry the organic layerwith MgSO₄. Filter off the drying agent and concentrate in vacuo toafford 17.07 g of 2-bromo-1-(5-chloro-2-methoxy-phenyl)-ethanone.

Dissolve 2-bromo-1-(5-chloro-2-methoxy-phenyl)-ethanone (17.07 g, 64.78mmol) in 150 mL of CHCl₃ and add hexamethylenetetramine (9.09 g, 64.84mmol) to the reaction mixture. Allow the reaction to stir at ambienttemperature for 23 hours. Collect the solids via filtration and wash thesolids with Et₂O. Slurry the solid in 100 mL of MeOH and add 50 mL ofconcentrated HCl to it dropwise. Heat the reaction to reflux and allowit to stir for 23 hours. Cool the reaction to ambient temperature andfilter off the solids. Slurry the filtrate in MeOH and again collect thesolids via filtration. Concentrate the filtrate to afford 10.99 g of2-amino-1-(5-chloro-2-methoxy-phenyl)-ethanone hydrochloride.

Dissolve 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride(8.0362 g, 41.92 mmol), 4-(dimethylamino)pryidine (1.0525 g, 8.62 mmol)and adipic acid monomethyl ester (6.71 g, 41.90 mmol) in 100 mL ofCH₂Cl₂ and allow it to stir at room temperature for 45 minutes. Addcrude 2-amino-1-(5-chloro-2-methoxy-phenyl)-ethanone hydrochloride(10.99 g, 46.55 mmol) and triethylamine (9.42 g, 93.1 mmol) to thereaction mixture and allow it to stir at ambient temperature for 21hours. Dilute the reaction with 200 mL of CH₂Cl₂ and wash it with 1 NHCl (2×), saturated aqueous NaHCO₃ (2×) and brine (1×). Dry the organiclayer with MgSO₄. Filter off the drying agent and concentrate in vacuoto afford 9.39 g of the crude titled product: mass spectrum: m/z=342.1(M+H).

Example 108 5-[5-(5-chloro-2-methoxy-phenyl)-oxazol-2-yl]-pentanoic acidmethyl ester

Dissolve5-[2-(5-Chloro-2-methoxy-phenyl)-2-oxo-ethylcarbamoyl]-pentanoic acidmethyl ester (9.39 g, 27.47 mmol) and 4-(dimethylamino)pyridine (6.54 g,53.53 mmol) in 250 mL of CH₂Cl₂ and cool the reaction mixture in an icebath. Add triphenylphosphinedibromide (22.54 g, 53.40 mmol) to thereaction portionwise over 15 minutes. Raise the reaction to ambienttemperature and allow it to stir for 17 hours. Wash the reaction withwater (1×) and brine (2×). Dry the organic layers with MgSO₄. Filter offthe drying agent and concentrate in vacuo to afford a crude residue.Purify the residue using silica gel chromotagraphy eluting withhexanes/ethyl acetate mixtures to afford 4.38 g of the titled product(49%): mass spectrum: m/z=324.1 (M+H).

Example 109 5-[5-(5-Chloro-2-hydroxy-phenyl)-oxazol-2-yl]-pentanoic acidmethyl ester

Dissolve 5-[5-(5-chloro-2-methoxy-phenyl)-oxazol-2-yl]-pentanoic acidmethyl ester (4.174 g, 12.89 mmol) in 50 mL of CH₂Cl₂ and cool thesolution to −78° C. Add boron tribromide (48.9 mL of a 1M solution inCH₂Cl₂) dropwise to the reaction and allow it to warm to ambienttemperature and allow it to stir for 27 hours. Cool the reaction to −78°C. and quench it by adding 100 mL of MeOH dropwise to the reactionmixture. Allow the reaction to warm to ambient temperature and stir for19 hours. Concentrate the reaction mixture in vacuo. Dissolve theresidue in CH₂Cl₂ and wash it with saturated aqueous NaHCO₃ and brine.Dry the organic layer with MgSO₄. Filter off the drying agent andconcentrate in vacuo to afford a crude residue. Purify the residue usingsilica gel chromotagraphy eluting with hexanes/ethyl acetate mixtures toafford 3.26 g of the titled product (82%): mass spectrum: m/z=310.09(M+H).

Example 110 5-[5-(5-Chloro-2-hydroxy-phenyl)-oxazol-2-yl]-pentanoic acid

Dissolve 5-[5-(5-chloro-2-hydroxy-phenyl)-oxazol-2-yl]-pentanoic acidmethyl ester (3.26 g, 10.52 mmol) in 50 mL of dioxane and add a solutionof LiOH (1.2671 g, 50.91 mmol) in 25 mL of water to it. Allow thereaction to stir at ambient temperature for 18 hours. Acidify thereaction with 5N HCl and concentrate the reaction mixture in vacuo toremove the dioxane. Extract the aqueous residue with EtOAc (3×). Dry thecombined organic layers with MgSO₄. Filter off the drying agent andconcentrate in vacuo to afford a crude residue. Triturate the residue incold EtOAc. Collect the product via filtration to afford 1.8837 g of thetitled product (60%): mass spectrum: m/z=296.1 (M+H).

Example 111 5-[4-(2-methoxy-phenyl)-thiazol-2-yl]-pentanoic acid methylester

Bubble gaseous ammonia through a gas dispersion tube through a rapidlystirred 0° C. solution of methyl adipoyl chloride (17.0 mL, 100.0 mmol)in dioxane (200 mL, anhydrous) for 30 minutes. Allow to warm to roomtemperature. After 1 hour, filter out solid ammonium chloride andconcentrate mother liquor to get 20 grams of a white solid. Add 20%i-PrOH/CHCl₃ to material, dry over MgSO₄ and concentrate to afford5-carbamoyl-pentanoic acid methyl ester (15.09 g, 95%).

Add THF (200 mL, anhydrous) to a sealed vessel containing5-carbamoyl-pentanoic acid methyl ester (11.94 g, 75.0 mmol) andphosphorous pentasulfide (16.67 g, 37.5 mmol). Flush vessel with N₂,seal, and sonicate for 75 minutes. Break up solids and stir sealed underN₂ at room temperature for 50 hours. Concentrate, triturate with boilingCHCl₃ and filter hot (×3). Concentrate combined mother liquors to get14.5 g yellow residue. Purify the residue by flash chromatography onsilica gel eluting with 0-60% EtOAc/hexanes to afford5-thiocarbamoyl-pentanoic acid methyl ester (7.72 g, 59%).

Add THF (125 mL, anhydrous) to a sealed vessel containing5-thiocarbamoyl-pentanoic acid methyl ester (7.25 g, 41.4 mmol) and2-bromo-2′-methoxyacetophenone (9.48 g, 41.4 mmol). Flush vessel withN₂, seal, and heat at 80° C. overnight. Cool to room temperature, addEtOAc, wash with saturated aqueous NaHCO₃ solution, brine, andbackextract from each aqueous layer with EtOAc. Dry combined organiclayers over MgSO₄ and concentrate to get 18 grams of purple residue.Adsorb on SiO₂ and purify the residue by flash chromatography on silicagel eluting with 0-20% EtOAc/hexanes to afford the title compound (9.7g, 77%). TLC (30% EtOAc/hexanes) R_(f)=0.38. MS (IS) 306 (M+1)⁺.

Example 112 5-[4-(2-methoxy-phenyl)-thiazol-2-yl]-pentanoic acid

Add a solution of LiOH.H₂O (2.62 g, 62.5 mmol) in water (60 mL) to arapidly stirred solution of5-[4-(2-methoxy-phenyl)-thiazol-2-yl]-pentanoic acid methyl ester (3.82g, 12.5 mmol) in dioxane (120 mL), stir at room temperature. After 1hour, acidify to pH 1 with 5N HCl solution and concentrate to remove themajority of the dioxane. Partition residue between 20% i-PrOH/CHCl₃ and1N HCl solution, separate layers. Backextract from aqueous layer with20% i-PrOH/CHCl₃ and dry combined organic layers over MgSO₄, andconcentrate to get 3 grams of a pink oil. Recrystallize from EtOAc toafford the title compound (2.52 g, 69%). MS (IS) 292 (M+1)⁺.

Example 113 5-[4-(2-hydroxy-phenyl)-thiazol-2-yl]-pentanoic acid

Dissolve 5-[4-(2-methoxy-phenyl)-thiazol-2-yl]-pentanoic acid methylester (3.20 g, 10.5 mmol) in acetic acid (50 mL, glacial), add HBr (50mL, 48% aqueous solution) and heat to reflux under N₂ for 6 hours. Addadditional HBr (20 mL, 48% aqueous solution) and heat at reflux under N₂overnight. Adjust to pH 4 with 5N NaOH solution, extract with EtOAc(×2), dry over MgSO₄ and concentrate to get 2.64 grams of a light brownsolid. Recrystallize from EtOAc/hexanes to afford the title compound(2.14 g, 74%). MS (IS) 278 (M+1)⁺.

Preparation 17 2-bromo-1-(2-hydroxy-4-methoxy-phenyl)-ethanone

Heat a suspension of copper (II) bromide (16.79 g, 75.2 mmol) in EtOAc(40 mL) to reflux under N₂. Add a solution of2′-hydroxy-4′-methoxyacetophenone (7.48 g, 45.0 mmol) in CHCl₃ (40 mL)to the suspension dropwise over 3 minutes. Attach a drying tube to thetop of the condenser and reflux for 6 hours. Cool to room temperatureand stir under N₂ overnight. Filter mixture and rinse filter cake withEtOAc and CHCl₃, concentrate mother liquor to get 12.75 grams of a greenoily solid. Adsorb on SiO₂ and purify the residue by flashchromatography on silica gel eluting with 0-20% EtOAc/hexanes to affordthe crude (approx. 75% pure) title compound (8.5 g, 77%).

Example 114 5-[4-(2-hydroxy-4-methoxy-phenyl)-thiazol-2-yl]-pentanoicacid methyl ester

Add THF (50 mL, anhydrous) to a roundbottom flask containing5-thiocarbamoyl-pentanoic acid methyl ester (2.63 g, 15.0 mmol) and2-bromo-1-(2-hydroxy-4-methoxy-phenyl)-ethanone (4.90 g, 15.0 mmol, 75%pure). Heat to reflux under N₂ overnight. Cool to room temperature, addEtOAc, wash with saturated NaHCO₃ solution, brine, and backextract fromeach aqueous layer with EtOAc. Dry combined organic layers over MgSO₄and concentrate to get 8 grams of a yellow solid. Adsorb on SiO₂ andpurify the residue by flash chromatography on silica gel eluting with0-20% EtOAc/hexanes followed by flash chromatography on silica geleluting with CHCl₃ to afford the title compound (2.69 g, 56%). MS (IS)322 (M+1)⁺.

Example 115 5-[4-(2-hydroxy-4-methoxy-phenyl)-thiazol-2-yl]-pentanoicacid

Add a solution of LiOH.H₂O (1.76 g, 41.9 mmol) in water (25 mL) to arapidly stirred solution of5-[4-(2-hydroxy-4-methoxy-phenyl)-thiazol-2-yl]-pentanoic acid methylester (2.69 g, 8.4 mmol) in dioxane (50 mL), stir at room temperature.After 1 hour, acidify to pH 1 with 5N HCl solution and cool under N₂ ina refrigerator. Filter out solids and rinse with ice cold water. Drysolids in a vacuum oven overnight to afford the title compound (2.50 g,96%). MS (IS) 308 (M+1)⁺.

Preparation 18 1-Ethynyl-2-methoxy-benzene

Add 2-iodo-anisole (17.55 g, 75.0 mmol), trimethylsilyl acetylene (15.9mL, 112.5 mmol), copper (I) iodide (0.29 g, 1.5 mmol), and THF (225 mL,anhydrous) to a dry round bottom flask. Add diisopropylamine (22.1 mL,157.5 mmol) and dichlorobis(triphenylphosphine) palladium (II) (1.58 g,2.3 mmol) and stir the mixture at room temperature under N₂. After 2.5hours, quench reaction with water and extract with EtOAc (×2). Washcombined organic layers with brine, dry over MgSO₄ and concentrate toget 20.8 grams of a black oil. Adsorb on SiO₂ and purify the residue byflash chromatography on silica gel eluting with 0-5% EtOAc/hexanes toafford 2-methoxy-phenylethynyl)-trimethylsilane (13.2 g, 86%).

Add a solution of potassium hydroxide (3.66 g, 65.2 mmol) in water (30mL) dropwise over 30 minutes to a stirred solution of(2-methoxy-phenylethynyl)-trimethylsilane (13.2 g, 64.6 mmol) inmethanol (275 mL) and stir at room temperature for 1.5 hours.Concentrate, add brine to residue, and extract with EtOAc. Dry organiclayer over MgSO₄ and concentrate to get 10.5 grams of a black oil.Adsorb on SiO₂ and purify the residue by flash chromatography on silicagel eluting with 0-5% EtOAc/hexanes to afford the title compound (7.6 g,89%).

Preparation 19 6-Nitro-hexanoic acid ethyl ester

Add silver nitrite (23.1 g, 150 mmol) to a stirred solution of ethyl6-bromo-hexanoate (17.7 mL, 100 mmol) in diethyl ether (125 mL,anhydrous) and heat to reflux under N₂ overnight. Filter through a padof Celite® and rinse pad with diethyl ether, concentrate to get 21 gramsof a yellow oil. Adsorb on SiO₂ and purify the residue by flashchromatography on silica gel eluting with 0-20% EtOAc/hexanes to affordthe title compound (14.0 g, 74%).

Example 116 5-[5-(2-Methoxy-phenyl)-isoxazol-3-yl]-pentanoic acid ethylester

Add 1,4-phenylenediisocyanate (14.5 g, 90.8 mmol) to a stirred solutionof 1-ethynyl-2-methoxy-benzene (4.0 g, 30.3 mmol) and 6-nitro-hexanoicacid ethyl ester (8.6 g, 45.4 mmol) in toluene (300 mL, anhydrous) andstir under N₂. Add triethylamine (12.7 mL, 90.8 mmol) and heat to refluxunder N₂. After 2.5 hours, filter mixture through a pad of Celite® andrinse with toluene. Concentrate to get 9 grams of a orange oil. Adsorbon SiO₂ and purify the residue by flash chromatography on silica geleluting with 0-20% EtOAc/hexanes to afford the title compound (14.0 g,74%).

Example 117 5-[5-(2-Methoxy-phenyl)-isoxazol-3-yl]-pentanoic acid

Add a solution of LiOH.H₂O (1.83 g, 43.5 mmol) in water (30 mL) to arapidly stirred solution of5-[5-(2-methoxy-phenyl)-isoxazol-3-yl]-pentanoic acid ethyl ester (2.64g, 8.7 mmol) in dioxane (60 mL) and stir overnight at room temperature.After 1 hour, acidify to pH 1 with 5N HCl solution and concentrate toremove the majority of the dioxane. Partition residue between 20%i-PrOH/CHCl₃ and 1N HCl solution and separate layers. Backextract fromaqueous layer with 20% i-PrOH/CHCl₃ and dry combined organic layers overMgSO₄, and concentrate to get 2.5 grams of a yellow oil. Recrystallizefrom EtOAc/hexanes to afford the title compound (1.96 g, 82%). MS (IS)276 (M+1)⁺.

Example 118 5-[5-(2-Hydroxy-phenyl)-isoxazol-3-yl]-pentanoic acid ethylester

Add boron tribromide (43 mL, 43 mmol, 1.0 M solution in CH₂Cl₂) dropwiseover 30 minutes to a stirred 0° C. solution of5-[5-(2-methoxy-phenyl)-isoxazol-3-yl]-pentanoic acid ethyl ester (5.20g, 17.1 mmol) in CH₂Cl₂ (45 mL). Allow to warm to room temperatureovernight. Add boron tribromide (17 mL, 17 mmol, 1.0 M solution inCH₂Cl₂) and stir at room temperature under N₂. After 4 hours, add borontribromide (17 mL, 17 mmol, 1.0 M solution in CH₂Cl₂) and stir at roomtemperature under N₂. After 2 hours, quench via dropwise addition ofethanol (50 mL, absolute). Concentrate, dissolve residue in CHCl₃, washwith saturated NaHCO₃ solution (×2), dry over MgSO₄ and concentrate toget 4.7 g of a tan solid. Adsorb on SiO₂ and purify the residue by flashchromatography on silica gel eluting with 3-20% EtOAc/hexanes to affordthe title compound (3.96 g, 80%). MS (IS) 290 (M+1)⁺.

Example 119 5-[5-(2-Hydroxy-phenyl)-isoxazol-3-yl]-pentanoic acid

Add a solution of LiOH.H₂O (2.87 g, 68.4 mmol) in water (60 mL) to arapidly stirred solution of5-[5-(2-hydroxy-phenyl)-isoxazol-3-yl]-pentanoic acid ethyl ester (3.96g, 13.7 mmol) in dioxane (120 mL) and stir at room temperatureovernight. After 1 hour, acidify to pH 1 with 5N HCl solution andconcentrate to remove the majority of the dioxane. Add water to residueand place in refrigerator overnight. Filter out solids, wash with water,dry in a 50° C. vacuum oven for 6 hours to afford the title compound(3.11 g, 87%). MS (IS) 262 (M+1)⁺.

Preparation 20 4-Chloro-2-ethynyl-1-methoxy-benzene

Add 4-chloro-2-iodo-anisole (10.9 mL, 75.0 mmol), trimethylsilylacetylene (15.9 mL, 112.5 mmol), copper (I) iodide (0.29 g, 1.5 mmol),and THF (225 mL, anhydrous) to a dry round bottom flask. Adddiisopropylamine (22.1 mL, 157.5 mmol) anddichlorobis(triphenylphosphine) palladium (II) (1.58 g, 2.3 mmol) andstir the mixture at room temperature under N₂ overnight. Quench reactionwith water and extract with EtOAc (×2). Wash combined organic layerswith brine, dry over MgSO₄ and concentrate to get 24 g of a black oil.Adsorb on SiO₂ and purify the residue by flash chromatography on silicagel eluting with 0-3% EtOAc/hexanes to afford(5-chloro-2-methoxy-phenylethynyl)-trimethylsilane (13.7 g, 76%).

Add a solution of potassium hydroxide (3.28 g, 58.5 mmol) in water (30mL) dropwise over 25 minutes to a stirred solution of(5-chloro-2-methoxy-phenylethynyl)-trimethylsilane (13.7 g, 57.4 mmol)in methanol (275 mL) and stir at room temperature for 2 hours.Concentrate, add brine to residue, and extract with EtOAc (×2). Dryorganic layer over MgSO₄ and concentrate to get 13 g of a black oil.Adsorb on SiO₂ and purify the residue by flash chromatography on silicagel eluting with 0-5% EtOAc/hexanes to afford the title compound (8.98g, 94%).

Example 120 5-[5-(5-chloro-2-methoxy-phenyl)-isoxazol-3-yl]-pentanoicacid ethyl ester

Add 1,4-phenylenediisocyanate (8.41 g, 52.5 mmol) to a stirred solutionof 4-chloro-2-ethynyl-1-methoxy-benzene (4.0 g, 30.3 mmol) and6-nitro-hexanoic acid ethyl ester (8.6 g, 45.4 mmol) in toluene (300 mL,anhydrous) and stir at room temperature under N₂. Add triethylamine (7.3mL, 52.5 mmol) and heat to reflux under N₂ overnight. Filter mixturethrough a pad of Celite® and rinse with toluene. Concentrate to get 6.6g of a orange oil. Adsorb on SiO₂ and purify the residue by flashchromatography on silica gel eluting with 0-40% EtOAc/hexanes to affordthe title compound (4.51 g, 76%). MS (IS) 338 (M+1)⁺.

Example 121 5-[5-(5-chloro-2-hydroxy-phenyl)-isoxazol-3-yl]-pentanoicacid ethyl ester

Add boron tribromide (39 mL, 39 mmol, 1.0 M solution in CH₂Cl₂) dropwiseto a stirred −78° C. solution of5-[5-(5-chloro-2-methoxy-phenyl)-isoxazol-3-yl]-pentanoic acid ethylester (4.39 g, 13 mmol) in CH₂Cl₂ (40 mL). Allow to warm to roomtemperature. After 2 hours, cool to −78° C. and add boron tribromide (13mL, 13 mmol, 1.0 M solution in CH₂Cl₂) dropwise and allow to warm toroom temperature overnight. Quench via dropwise addition of ethanol (60mL, absolute). Concentrate, dissolve residue in CHCl₃, wash withsaturated NaHCO₃ solution (×2), dry over MgSO₄ and concentrate to get3.7 g of a tan solid. Adsorb on SiO₂ and purify the residue by flashchromatography on silica gel eluting with 0-60% EtOAc/hexanes to affordthe title compound (3.3 g, 78%). MS (IS) 324 (M+1)⁺.

Example 122 5-[5-(5-chloro-2-hydroxy-phenyl)-isoxazol-3-yl]-pentanoicacid

Add a solution of LiOH.H₂O (2.14 g, 51.0 mmol) in water (30 mL) to arapidly stirred solution of5-[5-(5-chloro-2-hydroxy-phenyl)-isoxazol-3-yl]-pentanoic acid ethylester (3.30 g, 10.2 mmol) in dioxane (60 mL), stir at room temperatureovernight. After 1 hour, acidify to pH 1 with 5N HCl solution and placein refrigerator. Filter out solids and rinse with water to afford thetitle compound (2.73 g, 91%). MS (IS) 296 (M+1)⁺.

Example 123 5-[4-(2-amino-phenyl)-oxazol-2-yl]-pentanoic acid methylester

Dissolve methyl adipoyl chloride (27 mL, 159 mmol) in dioxane (300 mL)and place the vessel in a room temperature water bath. Carefully bubblein ammonia gas (excess) and allow the mixture to stir for 1-2 hours.Filter the mixture to remove solids. Suspend solids in CHCl₃ and filteragain. Concentrate the combined filtrates and dry in vacuo to give 24.87g (98%) of 5-carbamoyl-pentanoic acid methyl ester.

Combine 2-bromo-2′-nitroacetophenone (9.6 g, 39.3 mmol) with5-carbamoyl-pentanoic acid methyl ester (12.0 g, 75.5 mmol) and heat theneat mixture in a sealed vessel at 120-140° C. for about 6 hours. Coolthe mixture and add methanol and allow the mixture to stir overnight atroom temperature. Concentrate the mixture and partition the residuebetween aq NaHCO₃ and EtOAc. Dry the combined extracts over Na₂SO₄ andconcentrate. Initial chromatography over silica gel (CH₂Cl₂) followed bya second chromatography over silica gel (Hex/EtOAc) allowed for recoveryof 5-[4-(2-nitro-phenyl)-oxazol-2-yl]-pentanoic acid methyl ester (4.65g, 39%). MS (ES): (M+1)⁺ 305.1, 306.3 m/z.

Combine 5-[4-(2-nitro-phenyl)-oxazol-2-yl]-pentanoic acid methyl ester(2.5 g, 8.2 mmol) with 5% Pd/C (300 mg) and Pd/black (50 mg) in THF andreact with hydrogen (init. 39 psi) in a Parr® apparatus. When reductionis complete, filter the mixture through Celite® and concentrate thefiltrate. Chromatograph the residue over silica gel (MeOH/CH₂Cl₂) toallow for recovery of 2.05 g (91%) of5-[4-(2-amino-phenyl)-oxazol-2-yl]-pentanoic acid methyl ester. MS (ES):(M+1)⁺ 275.1, 276.2 m/z.

Example 124 5-[4-(2-Amino-phenyl)-oxazol-2-yl]-pentanoic acid

Combine 5-[4-(2-amino-phenyl)-oxazol-2-yl]-pentanoic acid methyl ester(2.5 g, 9.1 mmol) with THF (3 mL), EtOH (3 mL) and 1N NaOH (15 mL) andstir until hydrolysis is complete. Concentrate the mixture, dilute theresidue with water and adjust the pH to 2.5-3.5 with aq HCl. Extract themixture with EtOAc and dry the extracts over Na₂SO₄ beforeconcentrating. Chromatograph the residue over silica gel (EtOAc) toallow for recovery of 5-[4-(2-amino-phenyl)-oxazol-2-yl]-pentanoic acid(2.03 g, 86%). MS (ES): (M+1)⁺ 261.1 m/z.

Example 125 5-[4-(2-Methanesulfonylamino-phenyl)-oxazol-2-yl]-pentanoicacid methyl ester

Dissolve 5-[4-(2-amino-phenyl)-oxazol-2-yl]-pentanoic acid methyl ester(2.18 g, 7.96 mmol) in THF (50 mL) and add pyridine (1.20 mL, 14.8mmol). Add methanesulfonyl chloride (excess) and allow the mixture tostir at room temperature until reaction is complete. Concentrate themixture and quench the residue with ice/aq NaHCO₃ and extract withEtOAc. Dry the combined extracts over Na₂SO₄ and concentrate.Chromatograph the residue over silica gel (EtOAc/CH₂Cl₂) to allow forisolation of 5-[4-(2-methanesulfonylamino-phenyl)-oxazol-2-yl]-pentanoicacid methyl ester (2.8 g, 100%). MS (ES): (M+1)⁺ 353.2, 354.3 m/z.

Example 126 5-[4-(2-Methanesulfonylamino-phenyl)-oxazol-2-yl]-pentanoicacid

Combine 5-[4-(2-methanesulfonylamino-phenyl)-oxazol-2-yl]-pentanoic acidmethyl ester (2.75 g, 7.8 mmol) with THF (3 mL), EtOH (3 mL) and 1N NaOH(20 mL) and stir at room temperature until hydrolysis is complete.Concentrate the mixture and dilute the residue with water and adjust topH 3.0-3.5 with aq HCl. Extract the mixture with EtOAc and dry theextracts over Na₂SO₄ before concentrating. Chromatograph the residueover silica gel (EtOAc) to allow for recovery of5-[4-(2-methanesulfonylamino-phenyl)-oxazol-2-yl]-pentanoic acid (2.16g, 82%). MS (ES): (M+1)+339.2, 340.3.

Example 127 5-[4-(2-Acetylamino-phenyl)-oxazol-2-yl]-pentanoic acidmethyl ester

Dissolve 5-[4-(2-amino-phenyl)-oxazol-2-yl]-pentanoic acid methyl ester(4.05 g, 14.8 mmol) and triethylamine (2.26 mL, 16.2 mmol) in TI-IF (40mL) and stir at room temperature. Add acetyl chloride (1.16 mL, 16.2mmol) and allow the mixture to stir overnight at room temperature.Concentrate the mixture and partition the residue between aq NaHCO₃ andEtOAc. Dry the combined extracts over Na₂SO₄ before concentrating.Chromatograph the residue over silica gel (MeOH/CH₂Cl₂) which allows forthe isolation of 5-[4-(2-acetylamino-phenyl)-oxazol-2-yl]-pentanoic acidmethyl ester (3.51 g, 75%). MS (ES): (M+1)⁺ 317.2 m/z.

Example 128 5-[4-(2-Acetylamino-phenyl)-oxazol-2-yl]-pentanoic acid

Combine 5-[4-(2-acetylamino-phenyl)-oxazol-2-yl]-pentanoic acid methylester (3.26 g, 10.3 mmol) with THF (3 mL), EtOH (3 mL) and 1 N NaOH (40mL) and stir at room temperature until hydrolysis is complete.Concentrate the mixture and dilute the residue with water beforeadjusting to pH 3.5-4.0 with aq HCl. Extract the mixture with 1-2%MeOH/EtOAc and concentrate the combined extracts in vacuo. Chromatographthe resulting residue over silica gel (MeOH/CH₂Cl₂) which allows forisolation of 5-[4-(2-acetylamino-phenyl)-oxazol-2-yl]-pentanoic acid(2.76 g, 89%). MS (ES): (M−1)⁻ 301.2, 302.3.

Example 129 5-[5-(5-Chloro-2-hydroxy-phenyl)-isoxazol-3-yl]-pentanoicacid disodium salt

Add a solution of NaOH (739 mg, 18.5 mmol) in water (3 mL) to asuspension of 5-[5-(5-chloro-2-hydroxy-phenyl)-isoxazol-3-yl]-pentanoicacid (2.73 g, 9.23 mmol) in water (5 mL) and stir at room temperature.Heat to 55° C. for 1 hour, filter hot solution, concentrate, place in50-60° C. vacuum oven for 2 days. Scrape and crush solids, place in50-60° C. vacuum oven overnight to afford the title compound (3.14 g,100%). MS (IS) 296 (M+1)⁺.

Preparation 21 5-Nitro-pentanoic acid methyl ester

Add silver nitrite (29.84 g, 193.9 mmol) to a stirred solution of methyl5-bromovalerate (25.22 g, 129.3 mmol) in diethyl ether (165 mL,anhydrous) and heat to reflux under N₂ overnight. Filter through a padof Celite® and rinse pad with diethyl ether, concentrate to get 21 gyellow oil. Adsorb on SiO₂ and purify the residue by flashchromatography on silica gel eluting with 0-20% EtOAc/hexanes to afford7.87 g of the title compound (38%).

Example 130 4-[5-(2-Methoxy-phenyl)-isoxazol-3-yl]-butyric acid methylester

Add 1,4-phenylenediisocyanate (19.22 g, 120.0 mmol) to a stirredsolution of 1-ethynyl-2-methoxy-benzene (2204901, 5.29 g, 40.0 mmol) and5-nitro-pentanoic acid methyl ester (7.91 g, 49.1 mmol) in toluene (300mL, anhydrous) and stir under N₂. Add triethylamine (16.7 mL, 120.0mmol) and heat to reflux under N₂ overnight. Filter mixture through apad of Celite® and rinse with toluene. Concentrate to get 12.4 g yellowoil. Adsorb on SiO₂ and purify the residue by flash chromatography onsilica gel eluting with 0-30% EtOAc/hexanes to afford the title compound(14.0 g, 74%). MS (IS) 276 (M+1)⁺.

Example 131 4-[5-(2-methoxy-phenyl)-isoxazol-3-yl]-butyric acid

Add a solution of LiOH (0.98 g, 40.86 mmol) in water (30 mL) to arapidly stirred solution of4-[5-(2-Methoxy-phenyl)-isoxazol-3-yl]-butyric acid methyl ester (2.25g, 8.17 mmol) in dioxane (60 mL), stir at room temperature overnightQuench with 1N HCl solution and concentrate to remove the majority ofthe dioxane. Add 1N NaOH to adjust pH to 4-5, extract with EtOAc (×2).Dry combined organic layers over MgSO₄ and concentrate to get 2.15 gyellow oil. Recrystallize from EtOAc/hexanes to afford the titlecompound (1.67 g, 78%). MS (IS) 262 (M+1)⁺.

Example 132 4-[5-(2-methoxy-phenyl)-isoxazol-3-yl]-butyric acid methylester

Add boron tribromide (51.2 mL, 51.2 mmol, 1.0 M solution in CH₂Cl₂)dropwise over 30 minutes to a stirred −78° C. solution of5-[5-(2-methoxy-phenyl)-isoxazol-3-yl]-pentanoic acid ethyl ester (2.82g, 10.2 mmol) in CH₂Cl₂ (30 mL). Allow to warm to room temperatureovernight Cool to 0° C. and quench via dropwise addition of methanol (60mL, anhydrous). Warm to room temperature, wash with saturated NaHCO₃solution (×2), dry over MgSO₄ and concentrate to get 2.7 g yellow oil.Purify the residue by flash chromatography on silica gel eluting with5-50% EtOAc/hexanes to afford the title compound (2.28 g, 85%). MS (IS)262 (M+1)⁺.

Example 133 5-[5-(2-hydroxy-phenyl)-isoxazol-3-yl]-pentanoic acid

Add a solution of LiOH (1.04 g, 43.6 mmol) in water (15 mL) to a rapidlystirred solution of 4-[5-(2-methoxy-phenyl)-isoxazol-3-yl]-butyric acidmethyl ester (2.28 g, 8.7 mmol) in dioxane (120 mL), stir at roomtemperature overnight After 1 hour, acidify to pH 1 with 5N HCl solutionand concentrate to remove the majority of the dioxane. Add water toresidue and place in refrigerator overnight Filter out solids, wash withwater, dry in a 50° C. vacuum oven for 6 hours to afford the titlecompound (3.11 g, 87%). MS (IS) 262 (M+1)⁺.

Preparation 22 5-azido-pentanoic acid methyl ester

Add sodium azide (12.5 g, 193 mmol) to a rapidly stirred solution ofmethyl 5-bromovalerate (25.08 g, 129 mmol) in DMSO (200 mL, anhydrous).Stir at room temperature overnight under N₂. Add water (400 mL) and stirfor 30 minutes. Extract with Et₂O (×3) and wash combined Et₂O layerswith brine (×3). Dry organic layer over MgSO₄ and concentrate to givethe title compound (20.3 g, 100%).

Examples 134 and 1355-[4-(2-methoxy-phenyl)-[1,2,3]triazol-1-yl]-pentanoic acid methyl esterand 5-[5-(2-methoxy-phenyl)-[1,2,3]triazol-1-yl]-pentanoic acid methylester

Add 1-ethynyl-2-methoxy-benzene (2204901, 5.29 g, 40.0 mmol),5-azido-pentanoic acid methyl ester (9.43 g, 60.0 mmol), and toluene(120 mL, anhydrous) to a dry flask and heat at reflux under N2 overnightConcentrate and purify the residue by flash chromatography on silica geleluting with 0-50% EtOAc/hexanes then 0-25% EtOAc/toluene to afford thetitle compounds A (2.85 g) and B (2.99 g) in a combined 50% yield. NOESYexperiment confirmed the structure of A. MS (IS) 290 (M+1)⁺.

Example 136 5-[4-(2-hydroxy-phenyl)-[1,2,3]triazol-1-yl]-pentanoic acid

Combine 5-[4-(2-methoxy-phenyl)-[1,2,3]triazol-1-yl]-pentanoic acidmethyl ester (2.40 g, 8.29 mmol), hydrobromic acid (50 mL, 48% inwater), and acetic acid (25 mL, glacial) and heat to reflux under N₂ for4 hours. Add hydrobromic acid (25 mL, 48% in water) and acetic acid (25mL, glacial) and continue heating at reflux under N₂ overnight Cool toroom temperature, filter out solids and dry in a 50° C. vacuum ovenovernight to yield the title compound (1.45 g, 67%). MS (IS) 260 (M−1)⁻.

Example 137 5-[5-(2-hydroxy-phenyl)-[1,2,3]triazol-1-yl]-pentanoic acid

Combine 5-[5-(2-methoxy-phenyl)-[1,2,3]triazol-1-yl]-pentanoic acidmethyl ester (2.99 g, 10.3 mmol), hydrobromic acid (50 mL, 48% inwater), and acetic acid (50 mL, glacial) and heat to reflux under N₂ for3 hours. Add hydrobromic acid (20 mL, 48% in water) and continue heatingat reflux under N₂ overnight. Cool to room temperature, basify to pH 4with 5N NaOH solution. Extract from aqueous layer with EtOAc (×3), washcombined organic layers with brine, dry combined organic layers overMgSO₄ and concentrate to get 2.6 yellow oil. Concentrate, adsorb on SiO₂and purify the residue by flash chromatography on silica gel elutingwith 3-5% MeOH/CH₂Cl₂. Add Et₂O and concentrate (×5) in order to removesolvents and afford the title compound (2.16 g, 80%).

MS (IS) 262 (M+1)⁺.

Example 138 5-[5-(2-hydroxy-phenyl)-[1,2,3]triazol-1-yl]-pentanoic aciddisodium salt

Add a solution of NaOH (0.66 g, 16.53 mmol) in water (3 mL) to asuspension of 5-[5-(2-methoxy-phenyl)-[1,2,3]triazol-1-yl]-pentanoicacid (2.16 g, 8.27 mmol) and heat to 50° C. for 1 hour. Concentrate anddry in 50° C. vacuum oven overnight. Add Et₂O, sonicate and filter outsolids. Dry solids in 50° C. vacuum oven overnight (as solids are highlyhygroscopic) to afford the title compound (2.0 g, 79%). MS (IS) 262(M+1)⁺.

Preparation 23 N-(2-ethynyl-phenyl)-acetamide

Add copper (I) iodide (0.38 g, 2.0 mmol) to a stirred solution of2-iodo-aniline (21.9 g, 100.0 mmol) and trimethylsilyl acetylene (21.2mL, 150.0 mmol) in THF (300 mL, anhydrous) in a dry RB flask. Adddiisopropylamine (29.4 mL, 210.0 mmol) anddichlorobis(triphenylphosphine) palladium (II) (2.11 g, 3.0 mmol) andstir the mixture at room temperature under N₂. After 5 hours quenchreaction with water and extract with EtOAc (×3). Dry combined organiclayers over MgSO₄ and concentrate to get 27.5 g black oil. Adsorb onSiO₂ and purify the residue by flash chromatography on silica geleluting with 0-10% EtOAc/hexanes to afford2-trimethylsilanylethynyl-phenylamine (17.7 g, 93%). MS (IS) 190 (M+1)⁺.

Add acetic anhydride (2.6 mL, 27.5 mmol) dropwise to a stirred 0° C.solution of 2-trimethylsilanylethynyl-phenylamine (2238853, 4.73 g, 25.0mmol) in pyridine (100 mL) under N₂. Allow to warm to room temperatureovernight Concentrate and partition residue between EtOAc and 1N HCl,separate layers. Extract from aqueous layer with EtOAc, wash combinedorganic layers with brine, dry over MgSO₄ and concentrate. Adsorb onSiO₂ and purify the residue by flash chromatography on silica geleluting with 0-15% EtOAc/hexanes to affordN-(2-trimethylsilanylethynyl-phenyl)-acetamide (4.67 g, 81%). MS (IS)231.9 (M+1)⁺.

Add a solution of potassium hydroxide (1.62 g, 28.8 mmol) in water (100mL) dropwise to a rapidly stirred solution ofN-(2-trimethylsilanylethynyl-phenyl)-acetamide (4.44 g, 19.2 mmol) inmethanol (50 mL) and stir at room temperature for 4 hours. Concentrate,add brine to residue and extract with EtOAc (×3). Dry combined organiclayers over MgSO₄, concentrate, add CHCl₃ and concentrate to get 2.82 glight yellow solid. Purify the residue by flash chromatography on silicagel eluting with 0-30% EtOAc/hexanes to afford the title compound (2.56g, 84%). MS (IS) 160 (M+1)⁺.

Example 139 5-[5-(2-acetylamino-phenyl)-isoxazol-3-yl]-pentanoic acidethyl ester

Add 1,4-phenylenediisocyanate (5.15 g, 32.2 mmol) to a stirred solutionof N-(2-ethynyl-phenyl)-acetamide (2.56 g, 16.1 mmol) and6-nitro-hexanoic acid ethyl ester (4.56 g, 24.1 mmol) in toluene (200mL, anhydrous) and stir under N₂. Add triethylamine (4.5 mL, 32.2 mmol)and heat to reflux under N₂ overnight. Filter mixture through a pad ofCelite® and rinse with toluene. Concentrate to get 6.2 g yellow oil.Adsorb on SiO₂ and purify the residue by flash chromatography on silicagel eluting with 0-60% EtOAc/hexanes to afford the title compound (4.02g, 76%). Correct regioisomer is confirmed by NOESY. MS (IS) 331 (M+1)⁺.

Example 140 5-[5-(2-acetylamino-phenyl)-isoxazol-3-yl]-pentanoic acid

Add 2N HCl solution (100 mL, in water) to5-[5-(2-acetylamino-phenyl)-isoxazol-3-yl]-pentanoic acid ethyl ester(4.02 g, 12.2 mmol) and heat to reflux under N₂ overnight Cool to roomtemperature and concentrate to afford the title compound (3.46 g, 96%yield). MS (IS) 259 (M−1)⁻.

Preparation 245-{5-[4-methoxy-2-(2-trimethylsilanyl-ethoxymethoxy)-phenyl]-isoxazol-3-yl}-pentanoicacid ethyl ester

Add a fine suspension of iodine (25.4 g, 100 mmol) in CHCl₃ (700 mL)dropwise to a stirred mixture of 3-methoxyphenol (10.8 mL, 100 mmol),silver trifluoroacetate (22.1 g, 100 mmol), and CHCl₃ (100 mL) over 2 hunder N₂ at room temperature. Stir at room temperature under N₂ for 60 hthen filter mixture over Celite® and rinse pad with CHCl₃. Wash solutionwith aqueous 0.1 N Na₂S₂O₄ solution, saturated NaHCO₃ solution andbackextract from aqueous with CHCl₃. Dry combined organic layers overNa₂SO₄ and concentrate to get 26.6 g brown oil. Purify the residue byflash chromatography on silica gel eluting with CHCl₃ to afford2-iodo-5-methoxy-phenol (18.4 g, 74%). MS (IS) 249 (M−1)⁻.

Add copper (I) iodide (0.28 g, 1.5 mmol) to a stirred solution of2-iodo-5-methoxy-phenol (18.4 g, 73.6 mmol) and trimethylsilyl acetylene(15.6 mL, 110.4 mmol) in THF (225 mL, anhydrous) in a dry RB flask. Adddichlorobis(triphenylphosphine) palladium (II) (1.55 g, 2.2 mmol) anddiisopropylamine (21.7 mL, 154.5 mmol) and stir the mixture at roomtemperature under N₂ overnight Quench reaction with water and extractwith EtOAc (×3). Dry combined organic layers over MgSO₄ and concentrateto get 29 g black oil. Adsorb on SiO₂ and purify the residue by flashchromatography on silica gel eluting with 0-15% EtOAc/hexanes to afford5-methoxy-2-trimethylsilanylethynyl-phenol (11.5 g, 71%). MS (IS) 221(M+1)⁺.

Add diisopropylamine (11.0 mL, 78.3 mmol) to a rapidly stirred solutionof 5-methoxy-2-trimethylsilanylethynyl-phenol (11.5 g, 52.2 mmol) inCH₂Cl₂ at room temperature under N₂. Add 2-(trimethylsilyl)ethoxymethylchloride (13.9 mL, 78.3 mmol) dropwise over 5 minutes and stir at roomtemperature under N₂ overnight Acidify with aqueous 1N HCl solution, addwater, separate layers. Extract from aqueous layer with CH₂Cl₂ (×2), drycombined organic layers over MgSO₄ and concentrate. Adsorb on SiO₂ andpurify the residue by flash chromatography on silica gel eluting with0-5% EtOAc/hexanes to afford4-methoxy-2-(2-trimethylsilanyl-ethoxymethoxy)-1-trimethylsilanylethynyl-benzene(13.5 g, 74%). MS (IS) 351 (M+1)⁺.

Add a solution of potassium hydroxide (2.3 g, 40.4 mmol) in water (20mL) dropwise to a rapidly stirred solution of4-methoxy-2-(2-trimethylsilanyl-ethoxymethoxy)-1-trimethylsilanylethynyl-benzene(13.5 g, 38.5 mmol) in methanol (200 mL) and stir at room temperaturefor 1 hour. Concentrate, add brine to residue and extract with EtOAc(×2). Dry combined organic layers over MgSO₄ and concentrate to get 11.7g brown oil. Adsorb on SiO₂ and purify the residue by flashchromatography on silica gel eluting with 0-10% EtOAc/hexanes to afford[2-(2-ethynyl-5-methoxy-phenoxymethoxy)-ethyl]-trimethyl-silane (10.0 g,93%). MS (IS) 279 (M+1)⁺.

Add 1,4-phenylenediisocyanate (6.41 g, 40.0 mmol) to a stirred solutionof [2-(2-ethynyl-5-methoxy-phenoxymethoxy)-ethyl]-silane (5.57 g, 20.0mmol) and 6-nitro-hexanoic acid ethyl ester (7.56 g, 40.0 mmol) intoluene (150 mL, anhydrous) and stir under N₂. Add triethylamine (5.6mL, 40.0 mmol) and heat to reflux under N₂. After 2 hours add additionaltoluene (150 mL, anhydrous) and continue refluxing overnight. Filtermixture through a pad of Celite® and rinse with toluene. Concentrate toget 8.7 g yellow oil. Purify the residue by flash chromatography onsilica gel eluting with 0-20% EtOAc/hexanes then 0-30% Et₂O/hexanes toafford the title compound (5.20 g, 58%). Correct regioisomer isconfirmed by NOESY. MS (IS) 450 (M+1)⁺.

Example 141 5-[5-(2-hydroxy-4-methoxy-phenyl)-isoxazol-3-yl]-pentanoicacid ethyl ester

Add a solution of HCl (10 mL, conc.) in methanol (50 mL) to a rapidlystirred solution of5-[5-(2-Hydroxy-4-methoxy-phenyl)-isoxazol-3-yl]-pentanoic acid ethylester (4.68 g, 10.4 mmol) in THF (75 mL) and methanol (75 mL). Stir atroom temperature under N₂ overnight Quench reaction with saturatedNaHCO₃ solution and extract with EtOAc (×3). Dry combined organic layersover MgSO₄ and concentrate. Adsorb on SiO₂ and purify the residue byflash chromatography on silica gel eluting with 5-100% EtOAc/hexanes toafford the title compound (2.25 g, 71%). MS (IS) 306 (M+1)⁺.

Example 142 5-[5-(2-hydroxy-4-methoxy-phenyl)-isoxazol-3-yl]-pentanoicacid

Add a solution of LiOH (0.96 g, 40.0 mmol) in water (15 mL) to a rapidlystirred solution of5-[5-(2-hydroxy-4-methoxy-phenyl)-isoxazol-3-yl]-pentanoic acid ethylester (2.45 g, 8.0 mmol) in dioxane (25 mL), stir at room temperaturefor 2 hours. Acidify to pH 1 with 5N HCl solution affording a whiteprecipitate. Allow to cool to room temperature and add water. Filter outsolids, wash with water, dry in a 50° C. vacuum oven for 3 hours toafford the title compound (1.86 g, 80%). MS (IS) 292 (M+1)⁺.

Example 143 5-[4-(2-Hydroxy-phenyl)-oxazol-2-yl]-pentanoic acid disodiumsalt

Add a solution of NaOH (1.15 g, 28.75 mmol) in water (5 mL) to asuspension of 5-[4-(2-hydroxy-phenyl)-oxazol-2-yl]-pentanoic acid(compound of Example 6, 3.76 g, 14.38 mmol) in water (30 mL) and stir atroom temperature. Heat to 50° C. for 1 h, filter hot solution,concentrate, place in 50-60° C. vacuum oven overnight Scrape and crushsolids, place in 50-60° C. vacuum oven for 60 hours to afford the titlecompound (4.39 g, 100%). MS (IS) 260 (M−1)⁻.

Example 1445-{4-[2-(2,2,2-Trifluoro-acetylamino)-phenyl]-oxazol-2-yl}-pentanoicacid methyl ester

Dissolve 5-[4-(2-amino-phenyl)-oxazol-2-yl]-pentanoic acid methyl ester(1.4 g, 5.1 mmol) in Et₂O (25 mL) and add TEA (0.71 mL, 5.1 mmol). Coolthe mixture in an ice bath, add trifluoroacetic anhydride (0.72 mL, 5.1mmol) and allow the mixture to stir at 0° C. until reaction is complete.Quench the mixture with aq NaHCO₃ and extract with EtOAc. Dry thecombined extracts over Na₂SO₄ and concentrate. Chromatograph the residueover silica gel (CH₂Cl₂) to allow for isolation of5-{4-[2-(2,2,2-Trifluoro-acetylamino)-phenyl]-oxazol-2-yl}-pentanoicacid methyl ester (1.69 g, 89%). MS (ES): (M+1)⁺ 371.1, 372.3 m/z.

Example 1455-(4-{2-[Methyl-2,2,2-trifluoro-acetyl)-amino]-phenyl}-oxazol-2-yl)-pentanoicacid methyl ester

Dissolve5-{4-[2-(2,2,2-Trifluoro-acetylamino)-phenyl]-oxazol-2-yl}-pentanoicacid methyl ester (1.6 g, 4.3 mmol) in THF (25 mL) and stir at RT undernitrogen. Add sodium hydride in small portions (60%, 0.17 g, 4.3 mmol)and allow the mixture to stir until deprotonation is complete. Then addiodomethane (0.54 mL, 8.7 mmol) and heat at 50-60° C. until the reactionis complete. Concentrate the mixture and partition the residue betweenwater and EtOAc. Dry the combined extracts over Na₂SO₄ and concentrate.Chromatograph the residue over silica gel (3% EtOAc/CH₂Cl₂) to allow forisolation of5-(4-{2-[methyl-2,2,2-trifluoro-acetyl)-amino]-phenyl}-oxazol-2-yl)-pentanoicacid methyl ester (1.63 g, 98%). MS (ES): (M+1)⁺ 385.1, 386.3 m/z.

Example 146 5-[4-(2-Methylamino-phenyl)-oxazol-2-yl]-pentanoic acid

Combine5-(4-{2-[Methyl-2,2,2-trifluoro-acetyl)-amino]-phenyl}-oxazol-2-yl)-pentanoicacid methyl ester (2.38 g, 6.2 mmol) with THF (2 mL), EtOH (4 mL) and 2NNaOH (12 mL) and stir until hydrolysis is complete. Concentrate themixture, dilute the residue with water and adjust the pH to 4.0 with aqHCl. Extract the mixture with EtOAc and dry the extracts over Na₂SO₄before concentrating. Chromatograph the residue over silica gel (3%EtOAc/CH₂Cl₂) to allow for recovery of5-[4-(2-methylamino-phenyl)-oxazol-2-yl]-pentanoic acid (1.63 g, 96%).MS (ES): (M+1)⁺ 275.1 m/z.

Preparation 25 4-[4-(2-nitro-phenyl)-oxazol-2-yl]-butyric acid methylester

Dissolve methyl (4-chloroformyl) butyrate (20 mL, 144 mmol) in dioxane(250 mL) and place the vessel in a RT water bath. Carefully bubble inammonia gas (excess) and allow the mixture to stir for 1-2 h. Filter themixture to remove solids. Suspend solids in CHCl₃ and filter again.Concentrate the combined filtrates and dry in vacuo to give 19.7 g (94%)of 4-carbamoyl-butyric acid methyl ester. MS (ES): (M+Na)⁺ 168.1 m/z.

Combine 2-bromo-2′-nitroacetophenone (10.4 g, 42.6 mmol) with4-carbamoyl-butyric acid methyl ester (12.2 g, 84.1 mmol) and heat theneat mixture in a sealed vessel at 120-150° C. for about 4-6 h. Cool themixture, transfer to a round bottom flask with methanol and concentrate.Partition the residue between water and EtOAc. Dry the combined EtOAcextracts over Na₂SO₄ and concentrate. Initial chromatography over silicagel (EtOAc/CH₂Cl₂) followed by a second chromatography over silica gel(Hex/EtOAc) allowed for recovery of4-[4-(2-nitro-phenyl)-oxazol-2-yl]-butyric acid methyl ester (3.05 g,25%). MS (ES): (M+1)⁺ 291.1, 292.2 m/z.

Example 147 4-[4-(2-amino-phenyl)-oxazol-2-yl]-butyric acid methyl ester

Combine 4-[4-(2-nitro-phenyl)-oxazol-2-yl]-butyric acid methyl ester(3.0 g, 10.3 mmol) with Pd/C (5%, 450 mg) and Pd/black (50 mg) in THF(150 mL) and react with hydrogen (init. 40 psi) in a Parr® apparatus.When reduction is complete, filter the mixture through Celite® andconcentrate the filtrate. Drying allows for recovery of crude4-[4-(2-amino-phenyl)-oxazol-2-yl]-butyric acid methyl ester (2.36 g,88%). MS (ES): (M+1)⁺ 261.2 m/z.

Example 149 4-[4-(2-amino-phenyl)-oxazol-2-yl]-butyric acid

Combine 4-[4-(2-amino-phenyl)-oxazol-2-yl]-butyric acid methyl ester(2.36 g, 9.1 mmol) with THF (3 mL), EtOH (3 mL) and 2N NaOH (20 mL) andstir until hydrolysis is complete. Concentrate the mixture, dilute theresidue with water and adjust the pH to 3.5-4.0 with aq HCl. Extract themixture with EtOAc and dry the extracts over Na₂SO₄ beforeconcentrating. Chromatograph the residue over silica gel (MeOH/CH₂Cl₂)to allow for recovery of 4-[4-(2-amino-phenyl)-oxazol-2-yl]-butyric acid(1.63 g, 73%).

MS (ES): (M+1)⁺ 247.1 m/z.

Preparation 26 6-[4-(2-nitro-phenyl)-oxazol-2-yl]-hexanoic acid ethylester

Using a method similar to the procedure described for the preparation of4-carbamoyl-butyric acid methyl ester; 6-(chloroformyl) hexanoic acidethyl ester (10.7 g, 55.4 mmol) in dioxane (150 mL) and ammonia gas give6-carbamoyl-hexanoic acid ethyl ester (9.23 g, 96%). MS (ES): (M+1)⁺174.1 m/z.

Using a method similar to the procedure described for the preparation of4-[4-(2-nitro-phenyl)-oxazol-2-yl]-butyric acid methyl ester; reactionof 2-bromo-2′-nitroacetophenone (8.9 g, 36.5 mmol) with6-carbamoyl-hexanoic acid ethyl ester (9.1 g, 48.7 mmol) gives6-[4-(2-nitro-phenyl)-oxazol-2-yl]-hexanoic acid ethyl ester (3.19 g,26%). MS (ES): (M+1)⁺ 333.2, 334.4 m/z.

Example 150 6-[4-(2-amino-phenyl)-oxazol-2-yl]-hexanoic acid ethyl ester

Using a method similar to the procedure described for the preparation of4-[4-(2-amino-phenyl)-oxazol-2-yl]-butyric acid; reduction of6-[4-(2-nitro-phenyl)-oxazol-2-yl]-hexanoic acid ethyl ester (3.1 g, 9.3mmol) gives crude 6-[4-(2-amino-phenyl)-oxazol-2-yl]-hexanoic acid ethylester (2.8 g, 99%). MS (ES): (M+1)⁺ 303.3 m/z.

Example 151 6-[4-(2-amino-phenyl)-oxazol-2-yl]-hexanoic acid

Using a method similar to the procedure described for the preparation of4-[4-(2-amino-phenyl)-oxazol-2-yl]-butyric acid; hydrolysis of6-[4-(2-amino-phenyl)-oxazol-2-yl]-hexanoic acid ethyl ester (2.7 g, 8.9mmol) gives 6-[4-(2-amino-phenyl)-oxazol-2-yl]-hexanoic acid (2.03 g,83%). MS (ES): (M+1)⁺ 274.9 m/z.

Preparation 27 3-[4-(2-nitro-phenyl)-oxazol-2-yl]-propionic acid methylester

Using a method similar to the procedure described for the preparation of4-carbamoyl-butyric acid methyl ester; methyl-4-chloro-4-oxobutyrate (25mL, 204.5 mmol) in dioxane (250 mL) and ammonia gas gives succinamicacid methyl ester (24.5 g, 91%). MS (ES): (M+Na)⁺ 154.1 m/z.

Using a method similar to the procedure described for the preparation of4-[4-(2-nitro-phenyl)-oxazol-2-yl]-butyric acid methyl ester; reactionof 2-bromo-2′-nitroacetophenone (15.43 g, 63.2 mmol) with succinamicacid methyl ester (14.4 g, 110 mmol) gives3-[4-(2-nitro-phenyl)-oxazol-2-yl]-propionic acid methyl ester (6.24 g,36%). MS (ES): (M+1)^(+ 277.1) m/z.

Example 152 3-[4-(2-amino-phenyl)-oxazol-2-yl]-propionic acid methylester

Using a method similar to the procedure described for the preparation of4-[4-(2-amino-phenyl)-oxazol-2-yl]-butyric acid; reduction of3-[4-(2-nitro-phenyl)-oxazol-2-yl]-propionic acid methyl ester (6.2 g,9.3 mmol) in EtOAc gives 3-[4-(2-amino-phenyl)-oxazol-2-yl]-propionicacid methyl ester (2.9 g, 52%) after chromatography over silica gel(THF/hexanes). MS (ES): (M+1)⁺ 247.3 m/z.

Example 153 3-[4-(2-amino-phenyl)-oxazol-2-yl]-propionic acid

Using a method similar to the procedure described for the preparation of4-[4-(2-amino-phenyl)-oxazol-2-yl]-butyric acid; hydrolysis of3-[4-(2-amino-phenyl)-oxazol-2-yl]-propionic acid methyl ester (4.8 g,19.5 mmol) gives 3-[4-(2-amino-phenyl)-oxazol-2-yl]-propionic acid (3.84g, 85%) after chromatography over silica gel (MeOH/CH₂Cl₂).

MS (ES): (M+1)⁺ 233.1 m/z.

Preparation 28 2-Bromo-1-(5-chloro-2-methoxy-phenyl)-ethanone

Combine 5′-chloro-2′-hydroxyacetophenone (17.0 g, 100 mmol) withpotassium carbonate (15.1 g, 109 mmol) and iodomethane (12.5 mL, 200mmol) in DMF and stir in a sealed vessel overnight at RT. Remove DMF andpartition the residue between water and EtOAc. Dry the combined extractsover Na₂SO₄ and concentrate. Chromatograph the resulting residue oversilica gel (EtOAc/hexanes) to allow for isolation of1-(5-Chloro-2-methoxy-phenyl)-ethanone (17.1 g, 93%). MS (ES): (M+1)⁺185.1, 187.1 m/z.

Dissolve 1-(5-Chloro-2-methoxy-phenyl)-ethanone (8.5 g, 46.2 mmol) inCHCl₃ (40 mL) and add this mixture to a warmed slurry of CuBr₂ (20.6 g,92.4 mmol) in EtOAc (150 mL). Heat the resulting mixture near reflux forapprox. 3 h. Cool and filter the mixture and concentrate the resultingfiltrate. Chromatograph the resulting residue over silica gel (CH₂Cl₂)to allow for isolation of 2-bromo-1-(5-chloro-2-methoxy-phenyl)-ethanone(12.0 g, 99%). MS (ES): (M+1)^(+ 185.1, 187.1) m/z.

Example 154 5-[4-(5-Chloro-2-methoxy-phenyl)-oxazol-2-yl]-pentanoic acidmethyl ester

Combine 2-bromo-1-(5-chloro-2-methoxy-phenyl)-ethanone (12.0 g, 46 mmol)with 5-carbamoyl-pentanoic acid methyl ester (12.7 g, 79.9 mmol) andheat the neat mixture in a sealed vessel at 120-140° C. for about 6 h.Cool the mixture and add methanol and allow the mixture to stirovernight at room temperature. Concentrate the mixture and partition theresidue between aq NaHCO₃ and EtOAc. Dry the combined extracts overNa₂SO₄ and concentrate. Chromatography over silica gel (CHCl₃) allowsfor recovery of 5-[4-(5-chloro-2-methoxy-phenyl)-oxazol-2-yl]-pentanoicacid methyl ester (5.5 g, 37%).

MS (ES): (M+1)⁺ 324.1, 326.1 m/z.

Example 155 5-[4-(5-Chloro-2-hydroxy-phenyl)-oxazol-2-yl]-pentanoic acid

Dissolve 5-[4-(5-Chloro-2-methoxy-phenyl)-oxazol-2-yl]-pentanoic acidmethyl ester (0.32 g, 1.0 mmol) in CH₂Cl₂ (2 mL) under nitrogen at RT.Add boron tribromide (1M in CH₂Cl₂, 3 mL) and stir at RT until thereaction is complete. Quench the mixture into water and extract withEtOAc. Dry the combined extracts over Na₂SO₄ and concentrate.Chromatograph the resulting residue over silica gel (MeOH/CH₂Cl₂) toallow for isolation of5-[4-(5-chloro-2-hydroxy-phenyl)-oxazol-2-yl]-pentanoic acid (0.27 g,91%). MS (ES): (M+1)⁺ 296.1, 298.1 m/z.

Example 156 5-[4-(5-Chloro-2-hydroxy-phenyl)-oxazol-2-yl]-pentanoic aciddisodium salt

Combine 5-[4-(5-Chloro-2-hydroxy-phenyl)-oxazol-2-yl]-pentanoic acid(2.71 g, 9.2 mmol) with THF (10 mL), MeOH (10 mL) and 1N NaOH (18.4 mL)and stir at RT for 2 h. Concentrate the mixture and dry the residueovernight in vacuo at 40-50° C. to allow for recovery of5-[4-(5-Chloro-2-hydroxy-phenyl)-oxazol-2-yl]-pentanoic acid disodiumsalt (3.0 g, 96%). MS (ES): (M+1)⁺ 296.1, 298.1 m/z.

Preparation 29 2-Bromo-1-(2-methoxy-phenyl)-propan-1-one

Combine 2′-hydroxypropiophenone (4.55 mL, 33.3 mmol) with cesiumcarbonate (11 g, 33.7 mmol) and iodomethane (4.1 mL, 65.8 mmol) inacetone (100 mL) and stir in a sealed vessel at RT until the reaction iscomplete. Concentrate the mixture and partition residue between waterand EtOAc. Dry the combined extracts over Na₂SO₄ and concentrate.Chromatograph the resulting residue over silica gel (EtOAc/hexanes) toallow for isolation of 1-(2-methoxy-phenyl)-propan-1-one (5.01 g, 93%).MS (ES): (M+1)⁺ 165.1 m/z.

Dissolve 1-(2-methoxy-phenyl)-propan-1-one (5.0 g, 30.5 mmol) in CHCl₃(25 mL) and add this mixture to a warmed slurry of CuBr₂ (13.6 g, 60.1mmol) in EtOAc (75 mL). Heat the resulting mixture near reflux forapprox. 3 h. Cool and filter the mixture and concentrate the resultingfiltrate. Chromatograph the resulting residue over silica gel (CH₂Cl₂)to allow for isolation of 2-bromo-1-(2-methoxy-phenyl)-propan-1-one(7.32 g, 99%). MS (ES): (M+1)⁺ 243.0, 245.0 m/z.

Example 157 5-[4-(2-methoxy-phenyl)-5-methyl-oxazol-2-yl]-pentanoic acidmethyl ester

Combine 2-Bromo-1-(2-methoxy-phenyl)-propan-1-one (7.25 g, 29.8 mmol)with 5-carbamoyl-pentanoic acid methyl ester (9.5 g, 60.5 mmol) and heatthe neat mixture in a sealed vessel at 140-150° C. for about 4 h. Coolthe mixture, transfer to a flask using methanol and concentrate.Partition the residue between water and EtOAc. Dry the combined extractsover Na₂SO₄ and concentrate. Chromatography over silica gel(MeOH/CH₂Cl₂) allows for recovery of5-[4-(2-methoxy-phenyl)-5-methyl-oxazol-2-yl]-pentanoic acid methylester (3.63 g, 40%). MS (ES): (M+1)⁺ 304.2 m/z.

Example 158 5-[4-(2-hydroxy-phenyl)-5-methyl-oxazol-2-yl]-pentanoic acidmethyl ester

Dissolve 5-[4-(2-methoxy-phenyl)-5-methyl-oxazol-2-yl]-pentanoic acidmethyl ester (3.5 g, 11.5 mmol) in CH₂Cl₂ (50 mL) under nitrogen at RT.Add boron tribromide (1M in CH₂Cl₂, 29 mL) and stir at RT until thereaction is complete. Quench the mixture into ice/water and extract withCH₂Cl₂. Dry the combined extracts over Na₂SO₄ and concentrate.Chromatograph the resulting residue over silica gel (MeOH/CH₂Cl₂) toallow for isolation of5-[4-(2-hydroxy-phenyl)-5-methyl-oxazol-2-yl]-pentanoic acid methylester (2.61 g, 78%). MS (ES): (M+1)⁺ 290.2 m/z.

Example 159 5-[4-(2-hydroxy-phenyl)-5-methyl-oxazol-2-yl]-pentanoic acid

Combine 5-[4-(2-hydroxy-phenyl)-5-methyl-oxazol-2-yl]-pentanoic acidmethyl ester (2.9 g, 10.0 mmol) with THF (3 mL), EtOH (3 mL) and 2N NaOH(20 mL) and stir until hydrolysis is complete. Concentrate the mixture,dilute the residue with water and adjust the pH to 2.0-3.0 with aq HCl.Extract the mixture with EtOAc and dry the extracts over Na₂SO₄ beforeconcentrating. Chromatograph the residue over silica gel (MeOH/CH₂Cl₂)to allow for recovery of5-[4-(2-hydroxy-phenyl)-5-methyl-oxazol-2-yl]-pentanoic acid (2.14 g,78%). MS (ES): (M+1)⁺ 276.2 m/z.

Preparation 30 Mixture of 7-Bromo-6-oxo-heptanoic acid methyl ester and5-Bromo-6-oxo-heptanoic acid methyl ester

Dissolve 5-acetyl valeric acid (16.0 g, 111.1 mmol) in MeOH (200 mL) andwarm the mixture near 50° C. Add bromine (5.7 mL, 111.1 mmol) and heatthe mixture at reflux for 3 hours. Concentrate the mixture and dissolvethe resulting oil in Et₂O. Wash the organic solution with water andaqueous NaHCO₃ and dry over Na₂SO₄. Concentration gives the crudemixture of 7-bromo-6-oxo-heptanoic acid methyl ester and5-bromo-6-oxo-heptanoic acid methyl ester (25.3 g, 96%).

Preparation 31 2-Methoxy-thiobenzamide

Combine 2-methoxy benzamide (10.0 g, 66.2 mmol) with phosphoruspentasulfide (10.1 g, 22.7 mmol) and THF (200 mmol) in a sealed vesseland stir for 30 minutes at room temperature. Sonicate the mixture for 1hour and then allow the mixture to stand at room temperature. Decant theTHF solution from the solids and concentrate. Chromatograph theresulting residue over silica gel (MeOH/CH₂Cl₂) to give2-methoxy-thiobenzamide (7.05 g, 64%).

Example 160 5-[2-(2-methoxy-phenyl)-thiazol-4-yl]-pentanoic acid methylester

Combine 2-methoxy-thiobenzamide (3.5 g, 21 mmol) with the bromidemixture described in Preparation 30 (5.6 g, 23.7 mmol) in THF (100 mL)and heat the mixture near reflux for about 3 hours. Concentrate themixture in vacuo. Dissolve the residue in EtOAc, wash with water andaqueous NaHCO₃, and dry over Na₂SO₄. Concentrate the solution andchromatograph the residue over silica gel (EtOAc/hexanes) to recover5-[2-(2-methoxy-phenyl)-thiazol-4-yl]-pentanoic acid methyl ester (4.4g, 69%). MS (ES): (M+1)⁺ 306.2, 307.2 m/z.

Example 161 5-[2-(2-hydroxy-phenyl)-thiazol-4-yl]-pentanoic acid methylester

Dissolve 5-[2-(2-methoxy-phenyl)-thiazol-4-yl]-pentanoic acid methylester (3.3 g, 10.8 mmol) in CH₂Cl₂ (30 mL) under nitrogen at roomtemperature. Add boron tribromide (1M in CH₂Cl₂, 27 mL) and stir at roomtemperature until the reaction is complete. Quench the mixture withmethanol, add ice/water and extract with CH₂Cl₂. Dry the combinedextracts over Na₂SO₄ and concentrate. Chromatograph the resultingresidue over silica gel (EtOAc/CH₂Cl₂) to allow for isolation of5-[2-(2-hydroxy-phenyl)-thiazol-4-yl]-pentanoic acid methyl ester (2.2g, 70%). MS (ES): (M+1)⁺ 292.1 m/z.

Example 162 5-[2-(2-hydroxy-phenyl)-thiazol-4-yl]-pentanoic acid

Combine 5-[2-(2-hydroxy-phenyl)-thiazol-4-yl]-pentanoic acid methylester (2.2 g, 7.6 mmol) with THF (2.5 mL), EtOH (2.5 mL) and 2N NaOH (14mL) and stir until hydrolysis is complete. Concentrate the mixture,dilute the residue with water and adjust the pH to 2.0-3.0 using aqueousHCl. Extract the mixture with EtOAc and dry the extracts over Na₂SO₄before concentrating. Chromatograph the residue over silica gel(MeOH/CH₂Cl₂) to allow for recovery of5-[2-(2-hydroxy-phenyl)-thiazol-4-yl]-pentanoic acid (1.5 g, 72%). MS(ES): (M+1)⁺ 278.1 m/z.

Preparation 32 Mixture of 2-Methoxy-benzoic acid6-methoxycarbonyl-2-oxo-hexyl ester and 2-Methoxy-benzoic acid1-acetyl-4-methoxycarbonyl-butyl ester

Combine o-anisic acid (7.0 g, 46 mmol) with Na₂CO₃ (2.44 g, 23 mmol) inwater (50 mL) and heat at 50° C. until everything is in solution. Then,add the bromide mixture described in Preparation 30 (10.91 g, 46 mmol)in EtOH (100 mL) and heat the mixture near reflux for about 5 hours.After allowing the mixture to cool and stir overnight, concentrate themixture, add water, and extract with EtOAc. Dry the combined extractsover Na₂SO₄. Concentrate the solution and chromatograph the residue oversilica gel (EtOAc/hexanes) to recover a mixture of 2-methoxy-benzoicacid 6-methoxycarbonyl-2-oxo-hexyl ester and 2-methoxy-benzoic acidacetyl-4-methoxycarbonyl-butyl ester (13.0 g, 92%). MS (ES): (M+1)⁺309.1 m/z.

Examples 163 and 164 5-[2-(2-Methoxy-phenyl)-oxazol-4-yl]-pentanoic acidmethyl ester 4-[2-(2-methoxy-phenyl)-4-methyl-oxazol-5-yl]-butyric acidmethyl ester

Combine the mixture described in Preparation 32 (15 g, 48.7 mmol) withammonium acetate (11.25 g, 146 mmol) in HOAc (40 mL) and heat themixture at 120-140° C. for about 5 h. Cool the mixture and concentrateonce from MeOH and once from hexanes. Dilute the residue with water andextract with EtOAc. Wash the combined extracts with 3× aqueous NaHCO₃,and dry over Na₂SO₄. Concentrate the solution and chromatograph theresidue over silica gel (EtOAc/hexanes) to allow for separation of5-[2-(2-methoxy-phenyl)-oxazol-4-yl]-pentanoic acid methyl ester (1.0 g,7%, MS (ES): (M+1)⁺ 290.1 m/z) and4-[2-(2-methoxy-phenyl)-4-methyl-oxazol-5-yl]-butyric acid methyl ester(3.9 g, 28%), MS (ES): (M+1)⁺ 290.1 m/z.

Example 165 4-[2-(2-hydroxy-phenyl)-4-methyl-oxazol-5-yl]-butyric acidmethyl ester

Dissolve 4-[2-(2-methoxy-phenyl)-4-methyl-oxazol-5-yl]-butyric acidmethyl ester (2.1 g, 7.3 mmol) in CH₂Cl₂ (20 mL) under nitrogen at roomtemperature. Add boron tribromide (1M in CH₂Cl₂, 21.8 mL) and stir atroom temperature until the reaction is complete. Quench the mixture withmethanol, add water and extract with CH₂Cl₂. Dry the combined extractsover Na₂SO₄ and concentrate. Chromatograph the resulting residue oversilica gel (MeOH/CH₂Cl₂) to allow for isolation of4-[2-(2-hydroxy-phenyl)-4-methyl-oxazol-5-yl]-butyric acid methyl ester(1.55 g, 77%). MS (ES): (M+1)⁺ 276.1, 277.2 m/z.

Example 166 4-[2-(2-hydroxy-phenyl)-4-methyl-oxazol-5-yl]-butyric acid

Using a method similar to that described in Example 162, hydrolysis of4-[2-(2-hydroxy-phenyl)-4-methyl-oxazol-5-yl]-butyric acid methyl ester(2.75 g, 10 mmol) gives 2.31 g of4-[2-(2-hydroxy-phenyl)-4-methyl-oxazol-5-yl]-butyric acid (88%) afterpurification. MS (ES): (M+1)⁺ 262.0, 263.2 m/z

Example 167 5-[2-(2-hydroxy-phenyl)-oxazol-4-yl]-pentanoic acid methylester

Using a method similar to that described in Example 165,5-[2-(2-methoxy-phenyl)-oxazol-4-yl]-pentanoic acid methyl ester (1.65g, 5.7 mmol) with boron tribromide (1.0 M in CH₂Cl₂, 17.2 mL) in CH₂Cl₂gives 5-[2-(2-hydroxy-phenyl)-oxazol-4-yl]-pentanoic acid methyl ester(0.76 g, 48%). MS (ES): (M+1)⁺ 276.1, 277.2 m/z.

Example 168 5-[2-(2-hydroxy-phenyl)-oxazol-4-yl]-pentanoic acid

Using a method similar to that described in Example 162, hydrolysis of5-[2-(2-hydroxy-phenyl)-oxazol-4-yl]-pentanoic acid methyl ester (1.45g, 5.3 mmol) gives 5-[2-(2-hydroxy-phenyl)-oxazol-4-yl]-pentanoic acid(1.19 g, 86%) after purification.

MS (ES): (M+1)⁺ 262.1 m/z.

Preparation 33 Mixture of 7-(2-methoxy-benzoylamino)-6-oxo-heptanoicacid methyl ester 5-(2-methoxy-benzoylamino)-6-oxo-heptanoic acid methylester

Dissolve di-t-butyliminodicarboxylate (3.67 g, 16.9 mmol) in DMF (25 mL)under N₂ at RT and add NaH (60%, 0.75 g, 18.6 mmol). Stir the mixtureuntil deprotonation is complete and add the bromide mixture (4.0 g, 16.9mmol) described in Preparation 30 and heat the mixture overnight near50° C. Dilute the mixture with water and extract with EtOAc. Wash thecombined extracts with water and brine and dry over Na₂SO₄.Concentration and chromatography of the residue over silica gel(MeOH/CH₂Cl₂) gives a mixture of7-di-tert-butoxycarbonylamino-6-oxo-heptanoic acid methyl ester and5-di-tert-butoxycarbonylamino-6-oxo-heptanoic acid methyl ester (5.6 g,89%). MS (ES): (M+NH₄)^(+ 391.2) m/z.

Treat a mixture of 7-di-tert-butoxycarbonylamino-6-oxo-heptanoic acidmethyl ester and 5-di-tert-butoxycarbonylamino-6-oxo-heptanoic acidmethyl ester (7.5 g, 20 mmol) with Et₂O/HCl and stir the resultingmixture overnight at room temperature. Concentration and drying in vacuogives a mixture of 7-amino-6-oxo-heptanoic acid methyl ester and5-amino-6-oxo-heptanoic acid methyl ester hydrochlorides (4.1 g, 97%).MS (ES): (M+1)⁺ 174.1 m/z.

Combine a mixture of 7-amino-6-oxo-heptanoic acid methyl ester and5-amino-6-oxo-heptanoic acid methyl ester hydrochlorides (2.36 g, 11.3mmol) with o-anisic acid (1.71 g, 11.3 mmol),1-(dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.16 g, 11.3mmol), 1-hydroxybenzotriazole hydrate (11.3 mmol),N,N-diisopropylethylamine (5.88 mL, 33.8 mmol) in DMF and stir for 24-48hours. Dilute the mixture with water and extract with EtOAc. Wash thecombined extracts with brine and dry over Na₂SO₄ before concentrating.Chromatograph the residue over silica gel (MeOH/CH₂Cl₂) to allow forisolation of a mixture of 7-(2-methoxy-benzoylamino)-6-oxo-heptanoicacid methyl ester and 5-(2-methoxy-benzoylamino)-6-oxo-heptanoic acidmethyl ester (3.45 g, 99%). MS (ES): (M+1)⁺ 308.1 m/z.

Example 169 5-[2-(2-methoxy-phenyl)-oxazol-5-yl]-pentanoic acid methylester

Combine the mixture of Preparation 33 (2.9 g, 9.5 mmol) with phosphorusoxychloride (2.6 mL, 28.5 mmol) in DMF (15 mL) and heat the mixture atabout 90° C. for 1 hour. Cool the mixture, dilute with water and extractwith EtOAc. Wash the combined extracts with brine and dry over Na₂SO₄.Concentrate the solution and chromatograph the residue over silica gel(EtOAc/hexanes) to allow for separation of5-[2-(2-methoxy-phenyl)-oxazol-5-yl]-pentanoic acid methyl ester (0.96g, 35%). MS (ES): (M+1)⁺ 290.1 m/z.

Example 170 5-[2-(2-methoxy-phenyl)-oxazol-5-yl]-pentanoic acid

Combine 5-[2-(2-methoxy-phenyl)-oxazol-5-yl]-pentanoic acid methyl ester(2.1 g, 7.3 mmol) with THF (4 mL), EtOH (4 mL) and 2N NaOH (15 mL) andstir until hydrolysis is complete. Concentrate the mixture, dilute theresidue with water and adjust the pH to 3.0-4.0 using aq HCl. Extractthe mixture with CH₂Cl₂ and concentrate the extracts in vacuo. Dryinggives 5-[2-(2-methoxy-phenyl)-oxazol-5-yl]-pentanoic acid (1.8 g, 91%).MS (ES): (M+1)⁺ 274.1, 275.1 m/z.

Example 171 5-[2-(2-hydroxy-phenyl)-oxazol-5-yl]-pentanoic acid

Dissolve 5-[2-(2-methoxy-phenyl)-oxazol-5-yl]-pentanoic acid (1.75 g,6.4 mmol) in CH₂Cl₂ (20 mL) under nitrogen at room temperature. Addboron tribromide (1M in CH₂Cl₂, 15 mL) and stir at room temperatureuntil the reaction is complete. Cool the mixture, quench withwater/methanol (95/5) and extract with CH₂Cl₂. Dry the combined extractsover Na₂SO₄ and concentrate. Resubject the residue to hydrolysisconditions as described in Preparation 14. Chromatograph the resultingresidue over silica gel (MeOH/CH₂Cl₂) to allow for isolation of5-[2-(2-hydroxy-phenyl)-oxazol-5-yl]-pentanoic acid (1.45 g, 87%). MS(ES): (M+1)⁺ 262.1, 263.2 m/z.

Preparation 34 2-Bromo-1-(2-trifluoromethyl-phenyl)-ethanone

Dissolve 2′-(trifluoromethyl)-acetophenone (5.0 g, 26.6 mmol) in CHCl₃(25 mL) and add this mixture to a warmed slurry of CuBr₂ (11.86 g, 53.2mmol) in EtOAc (75 mL). Heat the resulting mixture near reflux forapprox. 5 hours. Cool and filter the mixture and concentrate theresulting filtrate. Chromatograph the resulting residue over silica gel(CH₂Cl₂) to allow for isolation of2-Bromo-1-(2-trifluoromethyl-phenyl)-ethanone (6.6 g, 93%).

Example 172 5-[4-(2-trifluoromethyl-phenyl)-oxazol-2-yl]-pentanoic acidmethyl ester

Combine 2-bromo-1-(2-trifluoromethyl-phenyl)-ethanone (6.6 g, 24.7 mmol)with 5-carbamoyl-pentanoic acid methyl ester (7.8 g, 49 mmol) and heatthe neat mixture in a sealed vessel at 140-150° C. for about 4.5 hours.Cool the mixture, dilute with water, and extract with EtOAc. Dry thecombined extracts over Na₂SO₄ and concentrate. Chromatography oversilica gel (CH₂Cl₂) allows for recovery of5-[4-(2-trifluoromethyl-phenyl)-oxazol-2-yl]-pentanoic acid methyl ester(3.47 g, 43%). MS (ES): (M+1)⁺ 328.2 m/z.

Example 173 5-[4-(2-trifluoromethyl-phenyl)-oxazol-2-yl]-pentanoic acid

Combine 5-[4-(2-trifluoromethyl-phenyl)-oxazol-2-yl]-pentanoic acidmethyl ester (3.35 g, 9.9 mmol) with THF (3 mL), EtOH (3 mL) and 2N NaOH(15 mL) and stir until hydrolysis is complete. Concentrate the mixture,dilute the residue with water and adjust the pH to 3.0-4.0 using aq HCl.Extract the mixture with EtOAc and dry the extracts over Na₂SO₄ beforeconcentrating. Chromatograph the residue over silica gel (MeOH/CH₂Cl₂)to allow for recovery of5-[4-(2-trifluoromethyl-phenyl)-oxazol-2-yl]-pentanoic acid (2.36 g,76%). MS (ES): (M+1)⁺ 314.2 m/z.

Formulation

Because the compound of formula II may contain a basic and/or acidicmoiety (i.e., amino and/or carboxylic acid), said compound may beformulated as a pharmaceutical salt, e.g., as the sodium orhydrochloride salt or as a salt described in “Handbook of PharmaceuticalSalts: Properties, Selection and Use”, Weinheim, N.Y.: VHCA; Wiley-VCH,2002. The compound of formula II is preferably formulated in a dosageunit form, i.e., in an individual delivery vehicle, for example, atablet or capsule, prior to administration to the recipient patient.Therefore, yet another embodiment of the present invention is apharmaceutical composition comprising a compound of formula II, or apharmaceutical salt thereof, an active agent, and a pharmaceuticalcarrier.

The present pharmaceutical compositions are prepared by known proceduresusing well-known and readily available ingredients. In making theformulations of the present invention, the delivery agent (formula IIcompound) will be mixed with an active agent and will usually be mixedwith a carrier, or diluted by a carrier, or enclosed within a carrierwhich may be in the form of a capsule, sachet, paper or other container.When the carrier serves as a diluent, it may be a solid, semisolid orliquid material which acts as a vehicle, excipient or medium for theactive ingredient.

Biological Assays Delivery Agent Formulation Development

For oral dosing of a GLP-1 compound, a pH range of 7.4 to 8.4 for eachformulation is typically employed, whereas for a MC4 agonist peptide, apH range of 6.8-7.2 (most typically 7.0) for the formulation of istypically utilized. A target delivery agent concentration of 150 mg/mLin both cases is also typical. Initial feasibility studies are conductedto determine final carrier formulations.

Briefly, 200 mg of delivery agent is weighed into a Type I glass vial,to which 1 mL of MilliQ water is added. Each mixture is visuallyinspected for solubility, followed by addition of NaOH to increasesolubility or HCl to decrease the pH to the oral dose range.Formulations are then diluted to 150 mg/mL with MilliQ water. Using thisapproach, the formulations generally fell into three categories: aqueoussoluble, nearly completely soluble (e.g., few undissolved particlesremaining, very fine aqueous suspensions or hazy suspensions), andaqueous insoluble (e.g., heavy suspensions). Delivery agents thatexhibited aqueous insolubility are formulated in 4% w/v (aqueous)hydroxypropylcellulose (Klucel® LF, Hercules, Wilmington, Del.) asneeded. In these cases, between 50 and 100 mg of agent is suspended inKlucel® LF in a Type I glass vial, to yield a concentration of 200mg/mL. For heavy aqueous and Klucel® LF suspensions, the preparationsare cooled on ice for 3 minutes, followed by probe sonication on ice for30 minutes using a Misonix Sonicator® Ultrasonic Processor XL (3/16^(th) inch microtip) to reduce particle size. Following pHadjustment with NaOH or HCl, the formulations are then diluted to 150mg/mL with MilliQ water or Klucel® LF.

Formulation of Stock Active Agent Solution

The GLP-1 compounds (e.g., Val⁸-Glu²²-GLP-1(7-37)OH andVal⁸-Glu²²-I³³-GLP-1(7-37)OH) and MC4 agonist peptides (e.g.,Ac-Arg-cyclo[Cys-Glu-His-D-Phe-Arg-Trp-Cys]-NH₂;Ac-cyclo[hCys-His-D-Phe-Arg-Trp-Cys]-NH₂;Ac-cyclo[hCys-His-D-Phe-Arg-Trp-penicillamine]—NH₂; andN-cyclohexanecarbonyl-cyclo[hCys-His-D-Phe-Arg-Trp-penicillamine]-NH₂)used herein are described in PCT Publication Number WO 03/072195 and PCTPatent Application No. PCT/US04/16625, filed June 17, respectively.

A stock solution of GLP-1 compound active agent is prepared as follows.Briefly, a known quantity of lyophilized active agent is weighed into aType I glass vial. MilliQ water is then added to yield an initialconcentration of about 7-10 mg/mL. Complete solubility of the peptide isachieved by slowly raising the pH of the medium to 10.5 with 1 N NaOHand 5 N NaOH, followed by incubation at room temperature for 30 minutes.A volume of 1 M Tris buffer, pH 8.0 is added to give a final bufferconcentration of 20 mM Tris, and the pH adjusted to pH 7.8 with 1N HCland 5 N HCl. The solution is then filtered through a low protein binding0.22 μM syringe filter (Millex GV, Millipore). The concentration of thepeptide filtrate is determined by UV spectroscopy (λ max=280 nm). Thesolution is then diluted to a stock concentration of about 5.0 mg/mLusing 20 mM Tris buffer, pH 7.8. The active agent solution is stored in1.0 mL aliquots at −70° C. until used.

A stock solution of MC4R agonist peptide is prepared as follows.Briefly, a known quantity of lyophilized MC4R agonist peptide is weighedinto a Type I glass vial. MilliQ water is then added to yield an initialconcentration of about 19-21 mg/mL. The pH is raised to 6.0 with 1 NNaOH and 5 N NaOH, followed by incubation at room temperature for 30minutes. The concentration of the peptide solution is determined by UVspectroscopy (max=280 nm; light scatter correction applied between 250nm and 410 nm). The solution is then stored as a stock, concentration ofabout 20.0 mg/mL. The peptide solution is stored, refrigerated 4-8° C.until used.

Rat Oral Delivery Method

Male Sprague-Dawley (femoral artery cannulated, Charles River,Wilmington, Mass.) rats weighing 250-300 g are used in these studies.Animals are housed in single house stainless steel cages and cared foraccording to Eli Lilly and Company Animal Care and Use Policies &Procedures. Animals are fasted for at least 12 hours (with free accessto water) before dose administration. Each experiment (deliveryagent+active agent) is conducted in a group of four rats. Finalformulations for each delivery agent are freshly prepared approximately5-10 minutes prior to in vivo dosing.

Specifically, delivery agent formulation (˜165 mg/mL stock) and GLP-1compound active agent solution (˜5.0 mg/mL stock) are added together toyield an admixture of delivery agent+active agent. The finalconcentrations in each such formulation are 150 mg/mL and 0.5 mg/mL,respectively. Formulations are dosed by oral gavage (PO) for a finaldose of 300 mg/kg delivery agent and 1.0 mg/kg active agent. One mL ofblood samples is collected in EDTA tubes from the systemic (femoralartery) cannula from each animal (one sample/time point) at 5, 10, and20 minutes. Tubes are chilled on ice immediately following collectionand centrifuged at approximately 5° C./3,000 rpm/15 minutes. Plasma isremoved, transferred into 12×75 mm polypropylene sample tubes with snapcaps, and stored immediately at −70° C. until analyzed by aradioimmunoassay.

In the case of an MC4 agonist peptide active agent, delivery agentformulation (˜165 mg/mL stock) and peptide solution (˜20.0 mg/mL stock)are added together to yield an admixture of delivery agent+active agent.The final concentrations in each such formulation are 150 mg/mL and 5.0mg/mL, respectively. Formulations are dosed by oral gavage (PO) for afinal dose of 300 mg/kg delivery agent and 10.0 mg/kg active agent. 0.40mL of blood sample is collected in heparin tubes from the systemic(femoral artery) cannula from each animal (one sample/time point) at, 5,15, 30, 60, 90 and 120 minutes. Tubes are chilled on ice immediatelyfollowing collection and centrifuged at approximately 5° C./3,000 rpm/15minutes. Plasma is removed, transferred into 96 well plates and storedimmediately at −70 C until analyzed by a LC/MS/MS.

Radioimmunoassay and Pharmacokinetic Analysis

Concentrations of immunoreactive active agent in rat plasma are assayedby a radioimmunoassay assay that non-specifically detects native peptideand metabolic products. These concentrations are subsequently used todetermine the reported pharmacokinetic parameters. Plasma samples aremixed with radiolabeled active agent and rabbit polyclonal antiserum andthen incubated overnight at ˜4° C. Bound and free forms ofimmunoreactive active agent are separated by precipitating the boundfraction by polyethylene glycol-assisted, secondary antibodyprecipitation. After collecting the bound fraction by centrifugation,the radioactivity is measured by a gamma counter. Data is analyzed by aweighted 4/5 parameter logistic algorithm. For GLP-1 compounds, thestandard curve ranges from 9.8 pg/mL to 10000 pg/mL and the upper andlower quantification limits are 150 pg/mL and 4000 pg/mL, respectively.For MC4 agonist peptides, the standard curve ranges from 5.0 ng/mL to5000 ng/mL and the upper and lower quantification limits are 10 ng/mLand 5000 ng/mL, respectively. Pharmacokinetic analysis is performedusing WinNonlin™ Version 3.0 (Pharsight Corporation, Mountain View,Calif.). Plasma concentration time data are reported as mean±standarddeviation (SD). Delivery agent efficiency is defined as area under theplasma concentration-time curve measured from 0 to 20 min (AUC) ofactive agent in the presence of each delivery agent. Representativecompounds of formula II (delivery agent) are tested with an active agentin the Rat Oral Delivery assay and the AUC of active agent in thepresence of delivery agent is greater than the AUC of the active agentin the absence of the delivery agent.

1-17. (canceled)
 18. A compound of formula (I), or a pharmaceuticallyacceptable salt thereof:

wherein R¹ is NH₂; R² is H; R³ is H; n is 2, 3, 4 or 5; and X is

where the phenyl substituent is attached at carbon atom number 4 and thealkanoic acid chain is attached at carbon atom number 2
 19. The compoundaccording to claim 18, selected from

and pharmaceutically acceptable salts thereof.
 20. A compound accordingto claim 19, which is:

or a pharmaceutical salt thereof.
 21. A compound according to claim 19,which is:

or a pharmaceutical salt thereof.
 22. A compound according to claim 19,which is:

or a pharmaceutical salt thereof.
 23. A compound according to claim 19,which is:

or a pharmaceutical salt thereof.
 24. A pharmaceutical compositioncomprising: a) a compound according to claim 18, or a pharmaceuticalsalt thereof; and b) a GLP-1 compound.
 25. The pharmaceuticalcomposition according to claim 24, wherein the GLP-1 compound is a GLP-1derivative.
 26. The pharmaceutical composition according to claim 24,wherein the GLP-1 compound is a GLP-1 analog.
 27. The compositionaccording claim 24, wherein the GLP-1 compound isVal⁸-Glu²²-GLP-1(7-37)OH.
 28. A pharmaceutical composition comprising:a) a compound of claim 18 or a pharmaceutical salt thereof; and b) anMC4 agonist peptide.
 29. The composition of claim 28, wherein the MC4agonist peptide is selected from the group consisting of:Ac-Arg-cyclo[Cys-Glu-His-D-Phe-Arg-Trp-Cys]-NH₂;Ac-cyclo[hCys-His-D-Phe-Arg-Trp-Cys]-NH₂;Ac-cyclo[hCys-His-D-Phe-Arg-Trp-penicillamine]-NH₂; andN-cyclohexanecarbonyl-cyclo[hCys-His-D-Phe-Arg-Trp-penicillamine]-NH₂.